Quinazoline derivatives for the treatment of viral infections and further diseases

ABSTRACT

This invention relates to quinazoline derivatives, processes for their preparation, pharmaceutical compositions, and their use in therapy of disorders in which the modulation of toll-like-receptors is involved.

This invention relates to quinazoline derivatives, processes for theirpreparation, pharmaceutical compositions, and their use in therapy.

The present invention relates to the use of quinazoline derivatives inthe treatment of viral infections, immune or inflammatory disorders,whereby the modulation, or agonism, of toll-like-receptors (TLRs) isinvolved. Toll-Like Receptors are primary transmembrane proteinscharacterized by an extracellular leucine rich domain and a cytoplasmicextension that contains a conserved region. The innate immune system canrecognize pathogen-associated molecular patterns via these TLRsexpressed on the cell surface of certain types of immune cells.Recognition of foreign pathogens activates the production of cytokinesand upregulation of co-stimulatory molecules on phagocytes. This leadsto the modulation of T cell behavior.

It has been estimated that most mammalian species have between ten andfifteen types of Toll-like receptors. Thirteen TLRs (named simply TLR1to TLR13) have been identified in humans and mice together, andequivalent forms of many of these have been found in other mammalianspecies. However, equivalents of certain TLR found in humans are notpresent in all mammals. For example, a gene coding for a proteinanalogous to TLR10 in humans is present in mice, but appears to havebeen damaged at some point in the past by a retrovirus. On the otherhand, mice express TLRs 11, 12, and 13, none of which are represented inhumans. Other mammals may express TLRs which are not found in humans.Other non-mammalian species may have TLRs distinct from mammals, asdemonstrated by TLR14, which is found in the Takifugu pufferfish. Thismay complicate the process of using experimental animals as models ofhuman innate immunity.

For detailed reviews on toll-like receptors see the following journalarticles. Hoffmann, J. A., Nature, 426, p 33-38, 2003; Akira, S.,Takeda, K., and Kaisho, T., Annual Rev. Immunology, 21, p 335-376, 2003;Ulevitch, R. J., Nature Reviews: Immunology, 4, p 512-520, 2004.

Compounds indicating activity on Toll-Like receptors have beenpreviously described such as purine derivatives in WO 2006 117670,adenine derivatives in WO 98/01448 and WO 99/28321, and pyrimidines inWO 2009/067081.

However, there exists a strong need for novel Toll-Like receptormodulators having preferred selectivity, higher potency, highermetabolic stability, and an improved safety profile compared to thecompounds of the prior art.

In the treatment of certain viral infections, regular injections ofinterferon (IFNα) can be administered, as is the case for hepatitis Cvirus (HCV), (Fried et. al. Peginterferon-alfa plus ribavirin forchronic hepatitis C virus infection, N Engl J Med 2002; 347: 975-82).Orally available small molecule IFN inducers offer the potentialadvantages of reduced immunogenicity and convenience of administration.Thus, novel IFN inducers are potentially effective new class of drugsfor treating virus infections. For an example in the literature of asmall molecule IFN inducer having antiviral effect see De Clercq, E.;Descamps, J.; De Somer, P. Science 1978, 200, 563-565.

IFNα is also given in combination with other drugs in the treatment ofcertain types of cancer (refer to Eur. J. Cancer 46, 2849-57, and CancerRes. 1992, 52, 1056 for examples). TLR 7/8 agonists are also of interestas vaccine adjuvants because of their ability to induce pronounced Th1response.

In accordance with the present invention a compound of formula (I) isprovided

or a pharmaceutically acceptable salt, solvate or polymorph thereof,wherein

R₁ is C₃₋₈alkyl, C₃₋₈alkoxy, C₂₋₆alkenyl or C₂₋₆alkynyl, each of whichis optionally substituted by one or more substituents independentlyselected from halogen, hydroxyl, amino, nitrile, ester, amide,C₁₋₃alkyl, C₁₋₃alkoxy or C₃₋₆cycloalkyl,

R₂ is hydrogen, halogen, hydroxyl, amine, C₁₋₇alkyl, C₁₋₇alkylamino,C₁₋₆alkoxy, (C₁₋₄)alkoxy-(C₁₋₄)alkyl, C₃₋₆cycloalkyl, C₄₋₇heterocycle,aromatic, bicyclic heterocycle, arylalkyl, heteroaryl, heteroarylalkyl,carboxylic amide, carboxylic ester each of which is optionallysubstituted by one or more substituents independently selected fromhalogen, hydroxyl, amino, C₁₋₆alkyl, di-(C₁₋₆)alkylamino,C₁₋₆alkylamino, C₁₋₆alkyl, C₁₋₆alkoxy, C₃₋₆cycloalkyl, carboxylic acid,carboxylic ester, carboxylic amide, heterocycle, aryl, alkenyl, alkynyl,arylalkyl, heteroaryl, heteroarylalkyl, or nitrile,

R₃ is hydrogen, halogen, hydroxyl, amine, C₁₋₇alkyl, C₁₋₇alkenyl,C₁₋₇alkynyl, C₁₋₇alkylamino, C₁₋₆alkoxy, (C₁₋₄)alkoxy-(C₁₋₄)alkyl,C₃₋₆cycloalkyl, C₄₋₇hetero-cycle, aromatic, bicyclic heterocycle,arylalkyl, heteroaryl, heteroarylalkyl, aryloxy, heteroaryloxy, ketone,nitrile each of which is optionally substituted by one or moresubstituents independently selected from halogen, hydroxyl, amino,C₁₋₆alkyl, di-(C₁₋₆)alkylamino, C₁₋₆alkylamino, C₁₋₆alkyl, C₁₋₆alkoxy,C₃₋₆cycloalkyl, carboxylic acid, carboxylic ester, carboxylic amide,heterocycle, aryl, alkenyl, alkynyl, arylalkyl, heteroaryl,heteroarylalkyl, or nitrile.

R₄ is hydrogen, halogen, hydroxyl, amine, C₁₋₇alkyl, C₁₋₇alkylamino,C₁₋₆alkoxy, (C₁₋₄)alkoxy-(C₁₋₄)alkyl, C₃₋₆cycloalkyl, C₄₋₇heterocycle,bicyclic heterocycle, arylalkyl, heteroarylalkyl, aryloxy, heteroaryloxyeach of which is optionally substituted by one or more substituentsindependently selected from halogen, hydroxyl, amino, C₁₋₆alkyl,di-(C₁₋₆)alkylamino, C₁₋₆alkylamino, C₁₋₆alkyl, C₁₋₆alkoxy,C₃₋₆cycloalkyl, carboxylic acid, carboxylic ester, carboxylic amide,heterocycle, aryl, alkenyl, alkynyl, arylalkyl, heteroaryl,heteroarylalkyl, or nitrile, and

R₅ is hydrogen, fluorine, chlorine or methyl with the proviso that

R₂, R₃, R₄, and R₅ cannot all be H.

In a first embodiment the present invention provides compounds offormula (I) wherein R₁ is butyl, pentyl or 2-pentyl and wherein R₂, R₃,R₄ and R₅ are as specified above.

In a further embodiment the current invention relates to compounds offormula (I) wherein R₁ is C₄₋₈ alkyl substituted with a hydroxyl, andwherein R₂, R₃, R₄ and R₅ are as specified above.

Another embodiment relates to compounds of formula (I) wherein R₁, whenbeing C₄₋₈alkyl substituted with hydroxyl, is one of the following:

In another embodiment the present invention provides compounds offormula (I) wherein R₅ is preferably hydrogen or fluorine and R₁, R₂,R₃, and R₄ are as described above.

The compounds of formula (I) and their pharmaceutically acceptable salt,solvate or polymorph thereof have activity as pharmaceuticals, inparticular as modulators of Toll-Like Receptor (especially TLR7 and/orTLR8) activity.

So, in a further aspect the present invention provides a pharmaceuticalcomposition comprising a compound of formula (I) or a pharmaceuticallyacceptable salt, solvate or polymorph thereof together with one or morepharmaceutically acceptable excipients, diluents or carriers.

Furthermore a compound of formula (I) or a pharmaceutically acceptablesalt, solvate or polymorph thereof according to the current invention,or a pharmaceutical composition comprising said compound of formula (I)or a pharmaceutically acceptable salt, solvate or polymorph thereof canbe used as a medicament.

Another aspect of the invention is that a compound of formula (I) or apharmaceutically acceptable salt, solvate or polymorph thereof, or saidpharmaceutical composition comprising said compound of formula (I) or apharmaceutically acceptable salt, solvate or polymorph thereof can beused accordingly in the treatment of a disorder in which the modulationof TLR7 and/or TLR8 is involved.

The term “alkyl” refers to a straight-chain or branched-chain saturatedaliphatic hydrocarbon containing the specified number of carbon atoms.

The term “halogen” refers to fluorine, chlorine, bromine or iodine.

The term “alkenyl” refers to an alkyl as defined above consisting of atleast two carbon atoms and at least one carbon-carbon double bond.

The term “alkynyl” refers to an alkyl as defined above consisting of atleast two carbon atoms and at least one carbon-carbon triple bond.

The term “cycloalkyl” refers to a carbocyclic ring containing thespecified number of carbon atoms.

The term “alkoxy” refers to an alkyl (carbon and hydrogen chain) groupsingular bonded to oxygen like for instance a methoxy group or ethoxygroup.

The term “aryl” means an aromatic ring structure optionally comprisingone or two heteroatoms selected from N, O and S, in particular from Nand O. Said aromatic ring structure may have 5, 6 or 7 ring atoms. Inparticular, said aromatic ring structure may have 5 or 6 ring atoms.

The term “aryloxy” refers to an aromatic ring structure. Said aromaticgroup is singularly bonded to oxygen, like for instance phenol.

The term “heteroaryloxy” refers to an aromatic ring structure optionallycomprising one or two heteroatoms selected from N, O and S. Saidaromatic group, containing 5 to 7 ring atoms, one of which is singularlybonded to oxygen like for instance hydroxypyridine.

The term “bicyclic heterocycle” means an aromatic ring structure, asdefined for the term “aryl” comprised of two fused aromatic rings. Eachring is optionally comprised of heteroatoms selected from N, O and S, inparticular from N and O.

The term arylalkyl” means an aromatic ring structure as defined for theterm “aryl” optionally substituted with an alkyl group.

The term “heteroarylalkyl” means an aromatic ring structure as definedfor the term “heteroaryl” optionally substituted by an alkyl group.

Heterocycle refers to molecules that are saturated or partiallysaturated and include ethyloxide, tetrahydrofuran, dioxane or othercyclic ethers. Heterocycles containing nitrogen include, for exampleazetidine, morpholine, piperidine, piperazine, pyrrolidine, and thelike. Other heterocycles include, for example, thiomorpholine,dioxolinyl, and cyclic sulfones.

Heteroaryl groups are heterocyclic groups which are aromatic in nature.These are monocyclic, bicyclic, or polycyclic containing one or moreheteroatoms selected from N, O or S. Heteroaryl groups can be, forexample, imidazolyl, isoxazolyl, furyl, oxazolyl, pyrrolyl, pyridonyl,pyridyl, pyridazinyl, pyrazinyl,

Pharmaceutically acceptable salts of the compounds of formula (I)include the acid addition and base salts thereof. Suitable acid additionsalts are formed from acids which form non-toxic salts. Suitable basesalts are formed from bases which form non-toxic salts.

The compounds of the invention may also exist in unsolvated and solvatedforms. The term “solvate” is used herein to describe a molecular complexcomprising the compound of the invention and one or morepharmaceutically acceptable solvent molecules, for example, ethanol.

The term “polymorph” refers to the ability of the compound of theinvention to exist in more than one form or crystal structure.

The compounds of the present invention may be administered ascrystalline or amorphous products. They may be obtained for example assolid plugs, powders, or films by methods such as precipitation,crystallization, freeze drying, spray drying, or evaporative drying.They may be administered alone or in combination with one or more othercompounds of the invention or in combination with one or more otherdrugs. Generally, they will be administered as a formulation inassociation with one or more pharmaceutically acceptable excipients. Theterm “excipient” is used herein to describe any ingredient other thanthe compound(s) of the invention. The choice of excipient dependslargely on factors such as the particular mode of administration, theeffect of the excipient on solubility and stability, and the nature ofthe dosage form.

The compounds of the present invention or any subgroup thereof may beformulated into various pharmaceutical forms for administrationpurposes. As appropriate compositions there may be cited allcompositions usually employed for systemically administering drugs. Toprepare the pharmaceutical compositions of this invention, an effectiveamount of the particular compound, optionally in addition salt form, asthe active ingredient is combined in intimate admixture with apharmaceutically acceptable carrier, which carrier may take a widevariety of forms depending on the form of preparation desired foradministration. These pharmaceutical compositions are desirably inunitary dosage form suitable, for example, for oral, rectal, orpercutaneous administration. For example, in preparing the compositionsin oral dosage form, any of the usual pharmaceutical media may beemployed such as, for example, water, glycols, oils, alcohols and thelike in the case of oral liquid preparations such as suspensions,syrups, elixirs, emulsions, and solutions; or solid carriers such asstarches, sugars, kaolin, diluents, lubricants, binders, disintegratingagents and the like in the case of powders, pills, capsules, andtablets. Because of their ease in administration, tablets and capsulesrepresent the most advantageous oral dosage unit forms, in which casesolid pharmaceutical carriers are obviously employed. Also included aresolid form preparations that can be converted, shortly before use, toliquid forms. In the compositions suitable for percutaneousadministration, the carrier optionally comprises a penetration enhancingagent and/or a suitable wetting agent, optionally combined with suitableadditives of any nature in minor proportions, which additives do notintroduce a significant deleterious effect on the skin. Said additivesmay facilitate the administration to the skin and/or may be helpful forpreparing the desired compositions. These compositions may beadministered in various ways, e.g., as a transdermal patch, as aspot-on, as an ointment. The compounds of the present invention may alsobe administered via inhalation or insufflation by means of methods andformulations employed in the art for administration via this way. Thus,in general the compounds of the present invention may be administered tothe lungs in the form of a solution, a suspension or a dry powder.

It is especially advantageous to formulate the aforementionedpharmaceutical compositions in unit dosage form for ease ofadministration and uniformity of dosage. Unit dosage form as used hereinrefers to physically discrete units suitable as unitary dosages, eachunit containing a predetermined quantity of active ingredient calculatedto produce the desired therapeutic effect in association with therequired pharmaceutical carrier. Examples of such unit dosage forms aretablets (including scored or coated tablets), capsules, pills, powderpackets, wafers, suppositories, injectable solutions or suspensions andthe like, and segregated multiples thereof.

Those of skill in the treatment of infectious diseases will be able todetermine the effective amount from the test results presentedhereinafter. In general it is contemplated that an effective dailyamount would be from 0.01 mg/kg to 50 mg/kg body weight, more preferablyfrom 0.1 mg/kg to 10 mg/kg body weight. It may be appropriate toadminister the required dose as two, three, four or more sub-doses atappropriate intervals throughout the day. Said sub-doses may beformulated as unit dosage forms, for example, containing 1 to 1000 mg,and in particular 5 to 200 mg of active ingredient per unit dosage form.

The exact dosage and frequency of administration depends on theparticular compound of formula (I) used, the particular condition beingtreated, the severity of the condition being treated, the age, weightand general physical condition of the particular patient as well asother medication the individual may be taking, as is well known to thoseskilled in the art. Furthermore, it is evident that the effective amountmay be lowered or increased depending on the response of the treatedsubject and/or depending on the evaluation of the physician prescribingthe compounds of the instant invention. The effective amount rangesmentioned above are therefore only guidelines and are not intended tolimit the scope or use of the invention to any extent.

Preparation of Compounds

Compounds of formula (I) are prepared according to scheme 1. The2,4-dichloroquinazolines can be reacted in separate steps to afford the2,4-diaminoquinazolines in acceptable yield. In the first step the2,4-dichloro-quinazoline is mixed or heated with an amine with orwithout a transition metal catalyst to afford the2-chloro-4-aminoquinazoline. After workup of the crude2-chloro-4-aminoquinazoline, the intermediate is heated in a pressurevessel with an ammonia source (for example, ammonia in methanol) andoptionally with CuO.

Compounds of formula (I) can also be prepared according to scheme 2.Substituted anthranilic esters (IV) were heated under acidic conditionsin the presence of excess cyanamide, using an alcoholic solvent (e.g.ethanol) or diglyme according to the method described in the literature(O'Hara et. al. JOC (1991) 56, p 776). Subsequent amine substitution ofthe 2-amino-4-hydroxyquinazolines (V) can proceed via several differentpathways. In one example, intermediates V can be heated in the presenceof phosphorous oxychloride (POCl₃) with or without solvent. Afterremoval of solvents, the amine can be added neat, or in the presence ofa polar solvent (e.g. acetonitrile) to afford VI at room temperature orby heating. A second approach is to react intermediates V with acoupling agent such as BOP or PyBOP in the presence of DBU and theamine. The reaction takes place in a polar solvent (e.g. DMF). A thirdmethod is to protect the 2-amino group in intermediate V with an acylgroup. Intermediate V is reacted with anhydride (e.g. acetic anhydride),typically at reflux for several hours. The solvents can be removed underreduced pressure and the crude can undergo subsequent reaction withPOCl₃ as described above. Facile removal of the protecting acyl group isdone via reaction in a basic solvent (e.g. sodium methoxide inmethanol).

EXPERIMENTAL SECTION Preparation of Intermediate A

To a mixture of 2,4-dichloro-6,7-dimethoxyquinazoline (500 mg, 1.9mmol), diisopropylethylamine (0.73 mL, 4.2 mmol), and acetonitrile (0.1mL) was added a solution of n-butylamine (0.19 mL, 1.9 mmol) inacetonitrile (5 mL) dropwise while stirring. The mixture was allowed tostir for one day at ambient temperature. Ethyl acetate was added, theorganic layer was washed with sat. aq ammonium chloride. The organiclayer was removed, dried over magnesium sulfate. The solids were removedvia filtration to afford crude A, used as such in the next step.

Preparation of Compound 1

Intermediate A (0.5 g, 1.7 mmol) was placed into a 20 mL pressure vesselwith 7N ammonia in methanol (15 mL) and to this was added CuO (242 mg,1.7 mmol). The vessel was sealed and the mixture was heated to 130° C.with stirring for 18 hours. The reaction was allowed to cool to roomtemperature. The solids were removed via filtration and the solvents ofthe filtrate were removed under reduced pressure. The crude material waspurified via reverse phase column chromatography (Vydac Denali C18column 10 μm, 250 g, 5 cm). Mobile phase (0.25% NH₄HCO₃ solution inwater, CH₃CN).

Preparation of 9

Step 1. Into a 500 mL round bottom flask equipped with a magnetic stirbar was placed methyl 2-amino-6-methoxybenzoate (25 g, 149.6 mmol),ethanol (200 mL), cyanamide (9.43 g, 224 mmol), and concentrated HCl (6mL). The mixture was allowed to stir at reflux for 6 hours. At one hourintervals, concentrated HCl (0.5 mL) was added. The reaction mixture wasallowed to cool to room temperature and the solid, V-1, was isolated viafiltration and washed with ethanol.

LC-MS m/z=192 (M+H).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 3.88 (s, 3H), 6.96 (dd, J=8.2, 3.1 Hz,2H), 7.69 (t, J=8.3 Hz, 1H), 8.28 (br. s., 2H), 12.67 (br. s., 1H)

Step 2.

Into a 50 mL vial was placed V-1 (250 mg, 1.24 mmol), anhydrous DMF (5mL), DBU (0.6 g, 3.73 mmol), and BOP (659 mg, 1.49 mmol). The mixturestirred at room temperature for 2 hours, n-butylamine (287 mg, 3.73mmol) was added and the reaction was allowed to stir at room temperaturefor 15 hours. The solvent was reduced in volume and the residue purifiedvia silica column chromatography using a dichloromethane to 10% methanolin dichloromethane gradient. The best fractions were pooled, thesolvents were removed under reduced pressure to afford 9.

The following intermediates were prepared according to the method toprepare V-1.

LC-MS m/z=240/242

¹H NMR (400 MHz, DMSO-d₆) δ ppm 3.09-3.55 (m, 2H), 7.09 (br. s., 1H),7.26 (dd, J=7.9, 1.3 Hz, 1H), 7.37-7.48 (m, 2H)

LC-MS m/z=196 (M+H)

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.00 (br. s., 2H) 7.13 (d, J=7.78 Hz,1H) 7.18 (d, J=8.28 Hz, 1H) 7.50 (t, J=8.03 Hz, 1H), phenol proton notobserved.

is LC-MS m/z=176 (M+H)

LC-MS m/z=180 (M+H)

¹H NMR (400 MHz, DMSO-d₆) δ ppm 6.98 (dd, J=11.0, 8.3 Hz, 1H), 7.13 (d,J=8.3 Hz, 1H), 7.51 (br. s., 2H), 7.64 (td, J=8.3, 5.8 Hz, 1H), 12.30(br. s, 1H)

LC-MS m/z=180 (M+H)

LC-MS m/z=239/241 (M+H)

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.32 (d, J=8.8 Hz, 1H), 7.49 (s, 2H),7.71 (br. s., 1H), 7.81 (dd, J=8.6, 2.4 Hz, 1H), 8.00 (d, J=2.4 Hz, 1H)

LC-MS m/z=192 (M+H)

LC-MS m/z=176 (M+H)

LC-MS m/z=180 (M+H)

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.01-7.16 (m, 2H), 7.56 (br. s., 2H),7.99 (t, J=7.7 Hz, 1H), 10.38-13.48 (m, 1H)

LC-MS m/z=196 (M+H)

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.41 (dd, J=8.5, 2.0 Hz, 1H), 7.55 (d,J=2.0 Hz, 1H), 7.98 (d, J=8.5 Hz, 1H), 8.49 (br. s., 2H), 10.79-13.69(m, 1H)

LC-MS m/z=176 (M+H)

¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.43 (s, 3H), 7.22 (d, J=1.0 Hz, 1H),7.24 (s, 1H), 7.89 (d, J=8.0 Hz, 1H), 8.29 (br. s., 2H), 12.65 (br. s,1H)

LC-MS m/z=192 (M+H)

LC-MS m/z=220 (M+H)

¹H NMR (400 MHz, DMSO-d₆) δ ppm 3.87-3.95 (m, 3H), 7.12-7.47 (m, 1H),7.83 (dd, J=8.3, 1.4 Hz, 1H), 7.99 (d, J=1.3 Hz, 1H), 8.07-8.13 (m, 1H),8.43 (br. s., 2H)

LC-MS m/z=198 (M+H)

LC-MS m/z=298 (M+H)

¹H NMR (400 MHz, DMSO-d₆) δ ppm 3.85 (s, 3H), 5.10 (s, 2H), 6.17 (br.s., 2H), 6.70 (s, 1H), 7.30-7.36 (m, 2H), 7.40 (t, J=7.4 Hz, 2H),7.44-7.48 (m, 2H), 10.82 (br. s., 1H)

LC-MS m/z=180 (M+H)

¹H NMR (400 MHz, DMSO-d₆) δ ppm 6.51-6.67 (m, 2H), 7.00-7.08 (m, 1H),7.42 (ddd, J=11.2, 7.9 1.3 Hz, 1H), 7.69 (dd, J=7.9, 0.6 Hz, 1H), 11.08(br. s., 1H)

LC-MS m/z=196 (M+H)

LC-MS m/z=176 (M+H)

¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.41 (s, 3H), 7.15 (t, J=7.5 Hz, 1H),7.43 (br. s., 2H), 7.55 (d, J=7.0 Hz, 1H), 7.80 (d, J=7.8 Hz, 1H),11.17-12.49 (m, 1H)

Preparation of 10

Step 1. Preparation of V-6. Into a 50 mL vial equipped with a magneticstir bar was placed V-3 (500 mg, 2.16 mmol), phenylboronic acid (342 mg,2.8 mmol), potassium carbonate (1.19 g, 8.62 mmol), dioxane (5.5 mL),water (1.8 mL), and tetrakis(triphenylphosphine)palladium (249 mg, 0.215mmol). Nitrogen gas was bubbled through the reaction mixture for 10minutes. The vial was sealed and heated to 130° C. The reaction cooledto room temperature and the solvents were removed under reducedpressure. The crude was purified via reverse phase column chromatography(RP Vydac Denali C18-10 μm, 200 g, 5 cm. Mobile phase 0.25% NH₄HCO₃solution in water, CH₃CN) to afford V-6.

LC-MS m/z=238 (M+H)

Step 2. Into a 50 mL vial equipped with a magnetic stir bar was placedV-6 (148 mg, 0.624 mmol), anhydrous DMF (3.5 mL), DBU (0.373 mL, 2.5mmol), BOP (345 mg, 0.78 mmol), then (S)-2-aminopentanol (322 mg, 3.12mmol). The reaction mixture was allowed to stir at room temperature for3 days. The volatiles were removed under reduced pressure and the crudewas partitioned between water and ethyl acetate. The organic layers werecombined, dried (magnesium sulfate), the solids were removed byfiltration, and the solvents of to the filtrate were removed underreduced pressure. The crude was purified via reverse phase columnchromatography (RP SunFire Prep C18 OBD-10 μm, 30×150 mm). Mobile phase(0.25% NH₄HCO₃ solution in water, CH₃CN) to afford 10.

Preparation of 11

Step 1. Into a 1 L round bottom flask equipped with a magnetic stir barwas placed V-1 (8.8 g, 46.03 mmol) and acetic anhydride (150 mL). Theflask was equipped with a reflux condenser and the mixture was heated toreflux with stirring for 15 hours. The precipitate was isolated byfiltration and washed with diisopropylether then dried in vacuo toafford a white solid, V-9.

LC-MS m/z=234 (M+H)

Step 2. Into a 250 mL round bottom flask equipped with a magnetic stirbar was added V-9 (4.5 g, 19.3 mmol), and acetonitrile (100 mL). POCl₃(5.56 mL, 59.8 mmol) was added dropwise over 30 minutes, followed by theaddition of DIPEA (10.3 mL, 59.8 mmol). The reaction mixture became abrown solution and stirred for 2 hours at room temperature. The reactionmixture was poured into 1M NaOH (100 mL) and extracted with ethylacetate (2×100 mL). The combined organic layers were dried over MgSO₄,the solids were removed via filtration and the filtrate was used as suchin the next step.

Step 3. The filtrate solution from step 2 in ethyl acetate was treatedwith DIPEA (9.2 mL, 53.6 mmol) and n-butylamine (3.5 mL, 35.8 mmol). Thereaction mixture was stirred for 16 hours at ambient temperature. Thesolvent was removed under reduced pressure and the crude wasreconstituted in dichloromethane and washed with water. The organiclayer was dried (MgSO₄), the solids were removed by filtration, and thesolvents of the filtrate were evaporated to dryness to obtain an orangesolid, V-11.

LC-MS m/z=289 (M+H)

Step 4. Into a 30 mL pressure tube was placed V-11 (2.8 g, 9.71 mmol),pyridine hydrochloride (6.73 g, 58.26 mmol), and pyridine (50 mL) andthe mixture was heated to 120° C. for 16 hours. The pyridine was removedunder reduced pressure. The crude was dissolved in a mixture ofdichloromethane/methanol: 95/5 and washed with a 1N HCl solution andwater. The organic layer was dried (MgSO₄), the solids were removed viafiltration and the solvents of the filtrate were removed under reducedpressure to afford VI-1.

LC-MS m/z=231 (M−H)

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.92 (t, J=7.37 Hz, 3H) 1.33-1.43 (m,2H) 1.50-1.59 (m, 2H) 3.41-3.49 (m, 2H) 5.79-5.88 (m, 1H) 6.02 (d,J=8.14 Hz, 1H) 6.91 (br. s., 2H) 6.99-7.04 (m, 1H) 10.78 (br. s., 1H)13.35 (br. s., 1H)

Step 5. Into a 100 mL flask was placed VI-1 (175 mg, 0.753 mmol), cesiumcarbonate (0.74 g, 2.26 mmol) and DMF (15 mL). The mixture was stirredat ambient temperature for 30 minutes. 2-bromoethyl methyl ether (0.089mL, 0.94 mmol) was added and the mixture was stirred for 16 hours atroom temperature. The solvent was removed under reduced pressure and thecrude residue was purified by HPLC (RP Vydac Denali C18-10 μm, 250 g, 5cm. Mobile phase (0.25% NH₄HCO₃ solution in water, methanol), the bestfractions were collected and the solvents were removed under reducedpressure to obtain 11 as a solid.

Preparation of 12

Step 1. V-2 was dissolved in DMF (15 mL) and purged with N₂ on an oilbath at 80° C. for 10 minutes. Thenbis(triphenylphosphine)palladium(II)dichloride (69 mg, 0.098 mmol),triphenylphosphine (57.6 mg, 0.22 mmol) and copper iodide (42.5 mg, 0.22mmol) were added. After 5 minutes of purging with N₂, diethylamine (3.15mL, 30.31 mmol) was added followed by the addition of 2-pyridylethyne(168 mg, 1.63 mmol). The vessel was closed and the reaction stirred at80° C. for 16 hours. The reaction mixture was poured into ice water, andthe precipitate was isolated by filtration, washed with water and driedunder vacuum. The product was stirred in dichloromethane for 30 minutes.The precipitate was isolated by filtration, washed with dichloromethaneand diisopropyl ether and dried under vacuo at 50° C. to obtain V-12.

LC-MS m/z=263 (M−H)

Step 2. To a solution of V-12 (300 mg, 1.15 mmol) in THF (50 mL) wasplaced 10% Pd/C (100 mg) under an N₂ (g) atmosphere. The reactionmixture stirred for 16 hours at room temperature, and subsequentlyfiltered over packed decalite. The solvent of the filtrate was removedunder reduced pressure to afford crude V-13, used as such in the nextstep.

LC-MS m/z=267 (M−H)

Step 3. Example 12 was prepared according to the method to prepare 9.

Preparation of 14

Step 1. Intermediates VI-2 and VI-3 was prepared according to the methodto prepare VI-1. VI-3 was isolated after stirring with diisopropyletherat room temperature.

VI-2: LC-MS m/z=337 (M+H)

VI-3: LC-MS m/z=379 (M+H)

Step 2. Compound 14 was prepared according to the method to prepareintermediate V-12.

Preparation of 15

Step 1. Into a 500 mL round bottom flask equipped with a magnetic stirbar was placed 3-aminophthalic acid hydrochloride (25 g, 115 mmol),ethanol (250 mL), cyanamide (7.25 g, 172 mmol), and concentrated HCl (6mL). The flask was equipped with a reflux condenser and the mixture wasallowed to stir at reflux for 6 hours. At one hour intervals,concentrated HCl (0.5 mL) was added via glass pipette. The reaction wasallowed to cool to room temperature, the solvents were removed underreduced pressure to afford a yellow oil. The crude was dried over silicagel then partially purified via silica gel column chromatography using adichloromethane to 10% methanol in dichloromethane gradient. The crude,yellow solid, V-14, was used as such in the next step.

LC-MS m/z=234 (M+H).

Step 2. Into a 100 mL round bottom flask equipped with a magnetic stirbar was placed V-14 (1.7 g, 7.29 mmol), anhydrous DMF (25 mL), DBU (3.3g, 21.87 mmol), and PyBOP (4.55 g, 8.75 mmol). The reaction mixture wasallowed to stir for 1 hour at room temperature. Then n-butylamine (2.1g, 29.2 mmol) was added and the mixture was allowed to stir for 15 hoursat room temperature. The solvent was removed under reduced pressure andthe crude was filtered through silica gel using 20% methanol indichloromethane. The solvents of the filtrate were removed under reducedpressure and the crude oil (15, 4 g) was purified via reverse phasecolumn chromatography (RP Vydac Denali C18-10 μm, 200 g, 5 cm). Mobilephase (0.25% NH₄HCO₃ solution in water, CH₃CN).

Preparation of 16

To a suspension of 10% Pd/C in methanol (25 mL) under a N₂ atmospherewas added compound 14 (111 mg, 0.39 mmol). The nitrogen atmosphere wasremoved and replaced by hydrogen gas. The mixture was allowed to stir atroom temperature until 2 equivalents of hydrogen gas were consumed. Thereaction mixture was filtered over packed decalite. The solvent of thefiltrate was removed under reduced pressure. The crude was purified viasilica gel column chromatography using a dichloromethane to 10% methanolin dichloromethane gradient to afford 16.

Preparation of 18

17 (625 mg, 2.28 mmol) was dissolved in anhydrous THF (10 mL). LAH (1Min THF, 3.42 mL, 3.42 mmol) was added dropwise and the reaction mixturewas stirred for 3 hours at room temperature. LC-MS showed completeconversion to the desired product. The reaction mixture was quenchedwith sat., aq. NH₄Cl, the solids were removed by filtration and thesolvents of the filtrate were removed under reduced pressure. Theresidue was purified via prep. HPLC yielding the product as a whitesolid.

Preparation of 19

A mixture of VI-2 (500 mg, 1.48 mmol),tetrakis(triphenylphosphine)palladium (86 mg, 0.074 mmol), and zinccyanide (106 mg, 0.89 mmol) in DMF (5 mL) in a 10 mL tube was placedunder microwave irradiation at 160° C. for 10 minutes. The mixture wascooled to room temperature and concentrated in vacuo. The residue waspartioned between water and dichloromethane. The organic layer wasseparated, dried (MgSO₄), the solvents were removed by filtration andthe solvents of the filtrate were concentrated in vacuo. The product wastriturated in CH₃CN, the solid was isolated by filtration. Acyldeprotection was afforded after treatment with sodium methoxide inmethanol at 60° C. for one hour. The mixture was cooled and the productprecipitated. The white solid, 19, was isolated by filtration and driedunder vacuum.

Preparation of 20

Into a 50 mL vial equipped with a magnetic stir bar and sparged withnitrogen gas was placed VI-3 (300 mg, 0.79 mmol), the boronic ester (198mg, 0.95 mmol), water (3 mL, degassed) and DME (6 mL, degassed), sodiumbicarbonate (199 mg, 2.37 mmol) and PdCl₂(PPh₃)₂ (55 mg, 0.079 mmol) wasadded and the mixture was heated to 90° C. for 1 hour. The mixture wascooled and ethyl acetate was added. The organic layer was separated,dried (MgSO₄), the solids were removed by filtration and the solvents ofthe filtrate were removed in vacuo. The residue was purified via silicagel column chromatography using a gradient of dichloromethane to 10%methanol in dichloromethane (containing ammonia). The product fractionswere collected and concentrated in vacuo. Acyl deprotection was affordedafter treatment with sodium methoxide in methanol at 60° C. for onehour. The solvents were removed under reduced pressure and the residuewas partitioned between water and dichloromethane. The organic layer wasseparated, dried (MgSO₄), the solvents were removed via filtration andthe solvents of the filtrate were removed in vacuo. The product wascrystallized from CH₃CN, isolated by filtration and dried in vacuo toobtain a white solid, 20.

Preparation of 21

In a first vial equipped with a magnetic stir bar and a screw capseptum, a solution of Pd₂(dba)₃ (6 mg, 0.007 mmol) and2-di-tert-butylphosphino-3,4,5,6-tetramethyl-2′,4′,6′-triisopropyI-1,1′-biphenyl (6 mg, 0.013 mmol) in toluene (0.5 mL) was flushed withN₂ gas then stirred at 120° C. for 3 minutes. A second vial, equippedwith a magnetic stir bar and a screw cap septum, was charged with2-methylimidazole (104 mg, 1.26 mmol) and K₃PO₄ (224 mg, 1.05 mmol),then VI-3 (200 mg, 0.53 mmol) and also flushed with N₂ (g). The premixedcatalyst solution followed by anhydrous toluene (0.5 mL) and t-butanol(1.0 mL) were added via syringe to the second vial (total 2 mL oftoluene: t-BuOH 1:1 solution). The reaction was heated to 120° C. for 12hours. The mixture was cooled and sodium methoxide (30% in methanol) wasadded. The mixture was heated at 60° C. for 1 hour. The mixture wascooled to room temperature and concentrated in vacuo. The residue waspartioned between water and dichloromethane. The organic layer wasseparated, dried (MgSO₄), the solids were removed by filtration and thesolvents of the filtrate were concentrated in vacuo. The crude waspurified by Prep HPLC (RP SunFire Prep C18 OBD-10 μm, 30×150 mm). Mobilephase (0.25% NH₄HCO₃ solution in water, CH₃CN). The product fractionswere collected and concentrated in vacuo to afford compound 21.

Preparation of 22

A mixture of VI-2 (500 mg, 1.48 mmol), tributyl(1-ethoxyvinyl)tin (0.626mL, 1.85 mmol), PdCl₂(PPh₃)₂ (220 mg, 0.31 mmol) in DMF (10 mL) washeated to 80° C. for 16 hours. The reaction mixture was cooled and HCl(1N, 2 mL) was added. The mixture was stirred at room temperature for 2hours then was poured into sat. aq. NaHCO₃ (100 mL) and the precipitatewas isolated by filtration, reconstituted in dichloromethane, dried(MgSO₄), the solids were removed by filtration and the solvents of thefiltrate were concentrated in vacuo. The product was purified via silicagel column chromatography using a gradient of dichloromethane to 5%methanol in dichloromethane, the product fractions were collected andconcentrated in vacuo. The product was triturated in DIPE, filtered anddried under vacuum to become a pale yellow solid. To the mixture wasadded methanol (6 mL) and sodium methoxide (0.716 mL) and was stirred at60° C. for 1 hour. The mixture was cooled and concentrated in vacuo. Theresidue was partioned between water and dichloromethane. The organiclayer was separated, dried (MgSO₄), the solids were removed byfiltration and solvents of the filtrate were concentrated in vacuo. Theproduct was triturated in DIPE, isolated by filtration and dried undervacuum to become a yellow solid, 22.

Preparation of 23

22 (59 mg, 0.23 mmol) was suspended in methanol (2 mL) and sodiumborohydride (9 mg, 0.23 mmol) was added. The mixture was stirred underN₂ (g) at room temperature for two hours. The mixture was diluted withdichloromethane (5 mL), then sat., aq. NH₄Cl (0.5 mL) was added followedby addition of NaHCO₃. The organic layer was dried (MgSO₄), the solidswere removed via filtration and the solvents of the filtrate wereconcentrated in vacuo. The product was triturated in DIPE, isolated byfiltration and dried under vacuum to become a pale yellow solid, 23.

Preparation of 24

Step 1. A 75 mL stainless steel autoclave was charged under nitrogenatmosphere with VI-2 (626 mg, 1.87 mmol), Pd(OAc)₂ (8 mg, 0.037 mmol),1,3-bis(diphenylphosphino)propane (31 mg, 0.074 mmol), potassium acetate(364 mg, 3.71 mmol), THF (20 mL), and methanol (20 mL). The autoclavewas closed and pressurized to 30 bar CO (g). The reaction mixture wasstirred for 16 hours at 120° C. The reaction mixture was allowed to coolto room temperature then concentrated in vacuo. The residue wasdissolved in water and extracted with dichloromethane. The organic layerwas dried (MgSO₄), the solids were removed by filtration and the solventof the filtrate was concentrated in vacuo. The product was purified on asilica column using a dichloromethane to 5% methanol in dichloromethanegradient. The product fractions were collected and concentrated in vacuoto obtain an off-white solid, VI-4.

Step 2. To a solution of VI-4 (190 mg, 0.69 mmol) in anhydrous THF (20mL) was added LAH (1M in THF, 1.04 mL, 1.04 mmol) at −75° C. under anitrogen atmosphere. The reaction was allowed to stir for two hourswhile it slowly warmed to 0° C. Then the mixture was cooled on aice-ethanol bath and carefully quenched by adding 15 mL ethyl acetatefollowed by Na₂SO₄ 10H₂O (2 g). The mixture was stirred for one hour andthen dried over MgSO₄, the solids were removed by filtration and thesolvent of the filtrate was removed under reduced pressure. The residuewas purified by prep. HPLC (RP Vydac Denali C18-10 μm, 200 g, 5 cm).Mobile phase (0.25% NH₄HCO₃ solution in water, CH₃CN), followed by SFCpurification

(Chiralpak Diacel AD 30×250 mm). Mobile phase (CO₂, methanol with 0.2%isopropylamine), the desired fractions were collected, and the solventswere removed under reduced pressure to afford 24.

Preparation of 25

Step 1. V-14 was reacted with trimethylacetylene according to the methodto prepare compound 14, to afford V-15.

LC-MS m/z=258 (M+H)

Step 2. VI-5 was prepared according to the method to prepare compound 9.Deprotection of the TMS group was performed in a NaHCO3, water, methanolmixture.

LC-MS m/z=357 (M+H)

Step 3. The hydrogenation was performed according to the method toprepare 16.

Preparation of 26

Step 1. Palladium catalyzed carbonylation of 2-bromo-4-isopropylanilinewas performed according to the procedure to prepare VI-4 with theexception that the reaction was run at 110° C. to afford2-amino-5-isopropylbenzoic acid.

LC-MS m/z=180 (M+H)

Step 2. V-16 was prepared according to the method to prepare V-1.

LC-MS m/z=204 (M+H)

Step 3. Example 26 was prepared according to the method to prepare 15.

Preparation of 27

to Step 1. Cyanamide was dissolved in ether and the mixture was stirredunder nitrogen gas. HCl (2M in ether) was added dropwise to the reactionmixture at ambient temperature and stirring continued for 2 hours atroom temperature. The precipitate, A-2 was isolated by filtration anddried in vacuo at 50° C.

Step 2. SO₂(CH₃)₂ (20.4 g, 217 mmol) was heated to melting. A-2 (3.3 g,29 mmol) was added and the resulting mixture was stirred and heated to120° C. to dissolve completely. Methyl5-(2-chloro-4-trifluoromethylphenoxy)-anthranilate (5 g, 14.5 mmol) wasadded in one part to the reaction mixture. Stirring was continued for 30minutes. The reaction mixture was treated with water (10 mL) and stirredfor 10 minutes. The precipitate, V-17, a white solid, was isolated byfiltration and dried in the vacuum oven.

LC-MS m/z=356 (M+H)

Step 3. Compound 27 was formed according to the method to prepare 15.

Preparation of 28

Step 1. A-3 (101 g, 0.44 mol) was dissolved in sulfuric acid (850 mL).This solution was cooled to 0° C. HNO₃ (18.3 mL, 0.44 mol) in sulfuricacid (200 mL) was added dropwise over 2 hours. The reaction mixture wasstirred for 45 minutes at −10° C., then poured into ice-water (6 L). Thesolvents were decanted and the residue was dissolved in dichloromethane(1.5 L). The aqueous layer was extracted with dichloromethane (1 L). Thecombined organic layers were dried (MgSO₄), the solids were removed byfiltration and the solvent was removed under reduced pressure to affordA-4, and the side product isomer A-5, separated via silica gel columnchromatography using a heptane to ethyl acetate gradient.

Step 2. Into a 500 mL erlenmeyer flask equipped with a magnetic stir barand sparged with nitrogen gas was placed methanol (100 mL, containing 2%thiophene), 5% Pt/C (2 g, 0.513 mmol) then placed under a hydrogenatmosphere. The reaction mixture was stirred for 16 hours at roomtemperature. The catalyst was removed by filtration and the volatiles ofthe filtrate were removed under reduced pressure. The residue waspurified on silica using a dichloromethane to dichloromethane:methanol9:1 gradient yielding a yellow oil, IV-2.

LC-MS m/z=244 (M+H)

Step 3. Intermediate V-18 was prepared according to the method toprepare V-17.

LC-MS m/z=254 (M+H)

Step 4. The procedure to prepare compound 9 was applied in the synthesisof 28 from V-18.

Preparation of compound 29

Step 1. Example 29 was afforded after catalytic hydrogenation of 27,according to the method described in the preparation of 25.

Preparation of 90

Step 1. 17 (12.515 g, 45.62 mmol) was dissolved in THF (100 mL). LiOH(3.83 g, 91.2 mmol) dissolved in water (20 mL) was added, followed bymethanol (50 mL). The reaction mixture was stirred overnight at roomtemperature. The volatiles were removed under reduced pressure, thesolid was washed with water and triturated with DIPE to afford VI-6 asoff-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.95 (t, J=7.4 Hz, 3H), 1.40 (dq,J=14.9, 7.3 Hz, 2H), 1.68 (quin, J=7.3 Hz, 2H), 3.54-3.65 (m, 2H),7.89-8.05 (m, 2H), 8.14-8.31 (m, 2H), 9.11 (br. s., 1H), 11.10 (br. s.,1H), 16.37 (br. s., 1H)

Step 2. Into a 50 mL vial was placed VI-6 (200 mg, 0.768 mmol), DMF (10mL), triethylamine (0.641 mL, 4.61 mmol), 3-aminopyridine (181 mg, 1.92mmol) and diethyl cyanophosphonate (0.233 mL, 1.54 mmol). The reactionwas allowed to stir for 2 hours at room temperature. The solvent wasremoved under reduced pressure and the crude was purified via reversephase column chromatography (Sunfire Prep C18, OBD 10 μm, 30×150 mm.Mobile phase (0.25% NH₄HCO₃ solution in water, methanol) to afford 90.

Synthetic Scheme for the Preparation of AA-9

Synthesis of Intermediate AA-3

To a solution of valeraldehyde (43 g, 500 mmol) in THF (1 L) was addedAA-2 (200 g, 532 mmol) and the reaction mixture was stirred for 16 hoursat room temperature. The solvents were evaporated and the residue wasdiluted in petroleum ether and filtered. The solvents of the filtratewere removed under reduced pressure and the residue was purified bysilica chromatography using a petroleum ether to 3% ethyl acetate inpetroleum ether gradient to give AA-3 (90 g) as a colorless oil.

¹H NMR (400 MHz, CDCl₃): δ ppm 6.81-6.77 (m, 1H), 5.68-5.64 (td, J=1.2Hz, 15.6 Hz, 1H), 2.11-2.09 (m, 2H), 1.406 (s, 9H), 1.38-1.26 (m, 4H),0.85-0.81 (t, J=7.2 Hz, 3H).

Synthesis of Compound AA-5

n-butyl lithium (290 mL, 725 mmol, 1.5 eq.) was added to a stirredsolution of AA-4 (165 g, 781 mmol) in THF (800 mL) at −78° C. Thereaction mixture was stirred for 30 minutes then AA-3 (90 g, 488.4 mmol)in THF (400 mL) was added and the reaction was stirred for 2 hours at−78° C. The mixture was quenched with sat., aq. NH₄Cl solution andwarmed to room temperature. The product was partitioned between ethylacetate and water. The organic phase was washed with brine, dried andevaporated. The residue was purified by column chromatography elutingwith 5% ethyl acetate in petroleum ether to afford a colorless oil, AA-5(132 g).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.36-7.16 (m, 10H), 3.75-3.70 (m, 2H),3.43-3.39 (d, J=15.2 Hz, 1H), 3.33-3.15 (m, 1H), 1.86-1.80 (m, 2H),1.47-1.37 (m, 2H), 1.32 (s, 9H), 1.26-1.17 (m, 7H), 0.83-0.79 (t, J=7.2Hz, 3H).

Synthesis of AA-6

AA-5 (130 g, 328 mmol) was dissolved in THF (1.5 L) and LAH (20 g, 526mmol) was added at 0° C. in small portions. The resulting mixture wasstirred at the same temperature for 2 hours and then allowed to warm toroom temperature. The mixture was quenched with a sat. aq. NH₄Clsolution. The product was partitioned between ethyl acetate and water.The organic phase was washed with brine, dried and evaporated. Thecombined organic layers were dried over sodium sulfate, the solids wereremoved via filtration and concentrated to afford crude AA-6 (100 g),which was used in the next step without further purification.

¹H NMR (400 MHz, CDCl₃): δ ppm 7.33-7.14 (m, 10H), 3.91-3.86 (m, 1H),3.80-3.77 (d, J=13.6 Hz, 1H), 3.63-3.60 (d, J=13.6 Hz, 1H), 3.43-3.42(m, 1H), 3.15-3.10 (m, 1H), 2.70-2.63 (m, 2H), 1.65-1.28 (m, 10H),0.89-0.81 (m, 3H).

Synthesis of AA-9

A solution of AA-6 (38 g, 116.75 mmol) and 10% Pd/C in methanol (200 mL)was hydrogenated under 50 PSI hydrogen at 50° C. for 24 hours. Thereaction mixture was filtered and the solvent was evaporated to givecrude product AA-7 (17 g).

The crude product was dissolved in dichloromethane (200 mL),triethylamine (26.17 g, 259.1 mmol) and di-tert-butyl dicarbonate (84.7g, 194.4 mmol) was added at 0° C. The resulting mixture was stirred atroom temperature for 16 hours. The mixture was partitioned betweendichloromethane and water. The organic phase was washed with brine,dried and evaporated. The residue was purified by silica gelchromatography eluting with 20% ethyl acetate in petroleum ether to giveAA-8 (13 g) as colorless oil.

¹H NMR (400 MHz, CDCl₃): δ ppm 4.08-4.03 (br, 1H), 3.68 (m, 1H),3.58-3.55 (m, 2H), 3.20-2.90 (br, 1H), 1.80-1.73 (m, 1H), 1.42-1.17 (m,15H), 0.85-0.82 (t, J=6.8 Hz, 3H).

AA-8 (42 g, 0.182 mol) was dissolved in dioxane (200 mL) and dioxane/HCl(4M, 200 mL) was added at 0° C. The resulting mixture was stirred atroom temperature for 2 h. The solvent was evaporated to afford the crudeproduct. A dichloromethane/petroleum ether mixture (50 mL, 1:1, v/v) wasadded to the crude product, and the supernatant was decanted. Thisprocedure was repeated two times to obtain an oil, AA-9 (26.6 g).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.04 (s, 3H), 3.60-3.49 (m, 2H),3.16-3.15 (m, 1H), 1.71-1.67 (m, 2H), 1.60-1.55 (m, 2H), 1.33-1.26 (m,4H), 0.90-0.87 (t, J=6.8 Hz, 3H).

Preparation of AA-10

AA-10 was prepared according to the preparation of AA-9, usingbutyraldehyde instead of valeraldehyde.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.07 (s, 3H), 4.85 (br, 1H), 3.57-3.45(m, 2H), 3.14-3.12 (m, 1H), 1.70-1.64 (m, 2H), 1.56-1.49 (m, 2H),1.38-1.30 (m, 2H), 0.90-0.80 (t, J=6.8 Hz, 3H).

TABLE 1 Compounds of formula (I). Method, Synthetic # STRUCTURE H NMR RtMethod  1

¹H NMR (360 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.3 Hz, 3 H), 1.31- 1.43(m, 2 H), 1.60 (t, J = 7.1 Hz, 2 H), 3.40-3.48 (m, 2 H), 3.79 (s, 3 H),3.79 (s, 3 H), 5.67 (s, 2 H), 6.63 (s, 1 H), 7.40 (s, 1 H), 7.44-7.50(m, 1 H) A, 0.67  2

¹H NMR (360 MHz, DMSO-d₆) δ ppm 0.85-0.93 (m, 3 H), 1.27- 1.37 (m, 4 H),1.57-1.68 (m, 2 H), 3.39-3.49 (m, 2 H), 3.78 (s, 3 H), 3.79 (s, 3 H),5.67 (s, 2 H), 6.63 (s, 1 H), 7.40 (s, 1 H), 7.47 (t, J = 5.7 Hz, 1 H)A, 0.86 Same method as to prepare 1.  3

¹H NMR (360 MHz, DMSO-d₆) δ ppm 0.79-0.91 (m, 3 H), 1.29 (m, J = 3.3 Hz,4 H), 1.59 (m, J = 6.6 Hz, 2 H), 1.64-1.70 (m, 1 H), 1.72-1.79 (m, 1 H),3.40-3.50 (m, 2 H), 3.80 (s, 3 H), 3.80 (s, 3 H), 4.33-4.43 (m, 1 H),4.48 (t, J = 5.1 Hz, 1 H), 5.68 (s, 2 H), 6.63 (s, 1 H), 7.09 (d, J =8.4 Hz, 1 H), 7.44 (s, 1 H) A, 0.74 Same method as to prepare 1.  4

¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 0.91 (t, J = 7.0 Hz, 3 H),1.28-1.48 (m, 5 H), 1.58-1.77 (m, 2 H), 3.48 (s, 1 H), 3.72 (dd, J =11.0, 6.3 Hz, 1 H), 3.88 (s, 3 H), 3.91 (s, 3 H), 4.34 (td, J = 6.8, 2.8Hz, 1 H), 4.78 (br. s., 2 H), 5.64 (d, J = 7.0 Hz, 1 H), 6.81 (s, 1 H),6.81 (s, 1 H) A, 0.68 Same method as to prepare 1.  5

¹H NMR (360 MHz, DMSO-d₆) δ ppm 0.88 (t, J = 7.3 Hz, 3 H), 1.23- 1.42(m, 2 H), 1.48-1.81 (m, 4 H), 3.39-3.48 (m, 2 H), 3.79 (s, 3 H), 3.80(s, 3 H), 4.38-4.46 (m, 1 H), 4.49 (t, J = 5.3 Hz, 1 H), 5.68 (s, 2 H),6.63 (s, 1 H), 7.08 (d, J = 8.4 Hz, 1 H), 7.44 (s, 1 H) A, 0.69 Samemethod as to prepare 1.  6

¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 0.95 (t, J = 7.3 Hz, 3 H),1.35-1.52 (m, 2 H), 1.60-1.71 (m, 2 H), 3.48 (s, 1 H), 3.71 (dd, J =11.0, 6.3 Hz, 1 H), 3.85 (s, 3 H), 3.85-3.88 (m, 1 H), 3.90 (s, 3 H),4.37 (td, J = 6.7, 3.3 Hz, 1 H), 4.85 (br. s., 2 H), 5.82 (d, J = 7.3Hz, 1 H), 6.78 (s, 1 H), 6.85 (s, 1 H) A, 0.69 Same method as toprepare 1.  7

¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 0.89- 0.96 (m, 4 H), 1.01 (d, J =1.0 Hz, 4 H), 1.25 (ddd, J = 13.7, 8.5, 7.4 Hz, 1 H), 1.47-1.65 (m, 1H), 1.77-1.92 (m, 1 H), 3.48 (s, 0 H), 3.81-3.84 (m, 1 H), 3.87 (s, 3H), 3.87 (s, 3 H), 4.21-4.31 (m, 1 H), 5.15 (br. s., 2 H), 6.04- 6.11(m, 1 H), 6.74 (s, 1 H), 6.86 (s, 1 H) Same method as to prepare 1.  8

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89-0.96 (m, 3 H), 1.31- 1.43 (m, 2 H),1.57-1.67 (m, 2 H), 3.44-3.52 (m, 2 H), 6.04 (s, 2 H), 7.01 (ddd, J =8.1, 7.0, 1.0 Hz, 1 H), 7.20 (dd, J = 8.4, 0.9 Hz, 1 H), 7.46 (ddd, J =8.3, 6.9, 1.4 Hz, 1 H), 7.75 (t, J = 5.4 Hz, 1 H), 7.98 (dd, J = 8.2,0.9 Hz, 1 H) A, 0.64 Same method as to prepare 1.  9

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.4 Hz, 3 H), 1.29- 1.44(m, 2 H), 1.63 (t, J = 7.3 Hz, 2 H), 3.55-3.64 (m, 2 H), 4.02 (s, 3 H),6.99 (dd, J = 8.3, 1.8 Hz, 2 H), 7.69 (t, J = 8.3 Hz, 1 H), 7.81- 8.29(m, 2 H), 9.10 (s, 1 H), 12.49 (s, 1 H) C, 0.83 10

¹H NMR (360 MHz, DMSO-d₆) δ ppm 0.80 (t, J = 1.00 Hz, 3 H) 0.83- 0.93(m, 1 H) 0.96-1.17 (m, 2 H) 1.20-1.35 (m, 1 H) 3.10- 3.26 (m, 2 H), 3.36(br. s., 2 H) 4.12 (td, J = 8.23, 4.39 Hz, 1 H) 4.56-4.74 (m, 1 H) 5.96(d, J = 8.42 Hz, 1 H) 7.18 (d, J = 1.00 Hz, 1 H) 7.37-7.64 (m, 6 H) 7.81(t, J = 1.00 Hz, 1 H) C, 0.88 11

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.28 Hz, 3 H) 1.36- 1.46(m, 2 H) 1.55-1.63 (m, 2 H) 3.37 (s, 3 H) 3.44 (td, J = 6.96, 5.14 Hz, 2H) 3.74-3.80 (m, 2 H) 4.24 (dd, J = 5.27, 3.76 Hz, 2 H) 6.04 (br. s, 2H) 6.57 (d, J = 7.53 Hz, 1 H) 6.77-6.81 (m, 1 H) 7.34 (t, J = 8.16 Hz, 1H) 7.97 (t, J = 5.02 Hz, 1 H) C, 0.85 See experimental section 12

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.90 (t, J = 7.37 Hz, 3 H) 1.32- 1.42(m, 2 H) 1.63-1.71 (m, 2 H) 3.05-3.12 (m, 2 H) 3.38- 3.48 (m, 2 H)3.52-3.59 (m, 2 H) 5.93 (s, 2 H) 6.88 (dd, J = 7.15, 1.21 Hz, 1 H) 7.07(dd, J = 8.25, 1.21 Hz, 1 H) 7.23-7.34 (m, 4 H) 7.71-7.76 (m, 1 H)8.53-8.56 (m, 1 H) C, 0.99 See experimental section 13

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.85-0.93 (m, 3 H), 1.25- 1.40 (m, 4 H),1.61 (t, J = 6.9 Hz, 2 H), 3.39-3.48 (m, 2 H), 6.13 (s, 2 H), 7.11 (d, J= 9.0 Hz, 1 H), 7.55 (dd, J = 8.8, 2.3 Hz, 1 H), 7.79- 7.90 (m, 1 H),8.25 (d, J = 2.3 Hz, 1 H) C, 0.99 Same method as to prepare 9 14

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.92 (t, J = 7.15 Hz, 3 H) 1.29- 1.45(m, 5 H) 1.51-1.67 (m, 2 H) 3.40-3.51 (m, 2 H) 4.60 (br. s., 1 H) 5.41(br. s., 1 H) 6.18 (br. s., 2 H) 7.11 (d, J = 8.58 Hz, 1 H) 7.41 (d, J =8.36 Hz, 1 H) 7.83- 7.96 (m, 1 H) 8.14 (br. s., 1 H) C, 0.74 Seeexperimental section 15

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.92 (m, J = 7.3, 7.3, 2.3 Hz, 6 H),1.29-1.45 (m, 4 H), 1.47- 1.60 (m, 4 H), 3.24-3.30 (m, 2 H), 3.39 (td, J= 6.8, 5.0 Hz, 2 H), 6.10 (s, 2 H), 6.96 (dd, J = 7.0, 1.3 Hz, 1 H),7.29 (dd, J = 8.4, 1.4 Hz, 1 H), 7.46 (t, J = 8.4 Hz, 1 H), 7.95 (t, J =4.8 Hz, 1 H), 8.88 (t, J = 5.6 Hz, 1 H) C, 0.97 See experimental section16

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.37 Hz, 3 H) 1.10 (d, J =6.16 Hz, 3 H) 1.30-1.42 (m, 2 H) 1.56-1.72 (m, 4 H) 2.53- 2.75 (m, 2 H)3.40-3.50 (m, 2 H) 3.57-3.66 (m, 1 H) 4.46 (d, J = 4.62 Hz, 1 H) 5.83(s, 2 H) 7.10 (d, J = 8.58 Hz, 1 H) 7.31 (dd, J = 8.58, 1.76 Hz, 1 H)7.65 (t, J = 5.39 Hz, 1 H) 7.76-7.84 (m, 1 H) C, 0.75 See experimentalsection 17

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.4 Hz, 3 H), 1.37 (dq, J =14.9, 7.4 Hz, 2 H), 1.66 (quin, J = 7.3 Hz, 2 H), 3.52-3.63 (m, 2 H),3.71 (br. s, 2 H), 3.93 (s, 3 H), 7.88 (dd, J = 8.5, 1.5 Hz, 1 H), 8.01(d, J = 1.5 Hz, 1 H), 8.46 (d, J = 8.5 Hz, 1 H), 9.67 (t, J = 5.4 Hz, 1H), 12.84 (s, 1 H) (HCl salt) C, 0.78 Same method as to prepare 9 fromV- 25 18

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.92 (t, J = 7.3 Hz, 3 H), 1.36 (dq, J =14.9, 7.4 Hz, 2 H) 1.60 (quin, J = 7.3 Hz, 2 H), 3.41-3.49 (m, 2 H),4.53 (s, 2 H), 65.24 (br. s., 1 H), 5.98 (s, 2 H), 6.96 (dd, J = 8.3,1.5 Hz, 1 H), 7.13 (s, 1 H), 7.69 (t, J = 5.4 Hz, 1 H), 7.92 (d, J = 8.5Hz, 1 H) C, 0.58 See experimental section 19

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.37 Hz, 3 H) 1.31- 1.44(m, 2 H) 1.55-1.65 (m, 2 H) 3.42-3.51 (m, 2 H) 6.57 (br. s., 2 H) 7.20(d, J = 8.80 Hz, 1 H) 7.71 (dd, J = 8.58, 1.76 Hz, 1 H) 8.02 (br. s., 1H) 8.55 (d, J = 1.76 Hz, 1 H) C, 0.83 See experimental section 20

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.40 Hz, 3 H) 1.33- 1.45(m, 2 H) 1.64 (m, J = 7.30, 7.30, 7.30, 7.30 Hz, 2 H) 3.41- 3.57 (m, 2H) 3.88 (s, 3 H) 5.93 (s, 2 H) 7.16 (d, J = 8.78 Hz, 1 H) 7.62-7.74 (m,2 H) 7.86 (s, 1 H) 8.04 (s, 1 H) 8.18 (d, J = 1.76 Hz, 1H) B, 4.24 Seeexperimental section 21

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.4 Hz, 3 H), 1.39 (dq, J =14.9, 7.4 Hz, 2 H), 1.59- 1.67 (m, 2 H), 2.18 (d, J = 0.9 Hz, 3 H), 3.48(td, J = 7.0, 5.6 Hz, 2 H), 6.11 (s, 2 H), 7.26 (d, J = 8.9 Hz, 1 H),7.39 (t, J = 1.2 Hz, 1 H), 7.71 (dd, J = 9.0, 2.5 Hz, 1 H), 7.78 (t, J =5.4 Hz, 1 H), 8.05 (d, J = 1.5 Hz, 1 H), 8.18 (d, J = 2.3 Hz, 1 H) B,4.5 see experimental section 22

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.26 Hz, 3 H) 1.26- 1.49(m, 2 H) 1.64 (quin, J = 7.21 Hz, 2 H) 2.58 (s, 3 H) 3.50 (q, J = 6.53Hz, 2 H) 6.43 (br. s., 2 H) 7.17 (d, J = 8.80 Hz, 1 H) 7.96 (d, J = 8.80Hz, 1 H) 8.19 (br. s., 1 H) 8.67 (s, 1 H) C, 0.73 see experimentalsection 23

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.37 Hz, 3 H), 1.30- 1.42(m, 5 H) 1.61 (quin, J = 7.32 Hz, 2 H), 3.42-3.50 (m, 2 H) 4.70- 4.77(m, 1 H) 5.07-5.16 (m, 1 H) 5.93 (s, 2 H) 7.15 (d, J = 8.36 Hz, 1 H),7.48 (dd, J = 8.58, 1.54 Hz, 1 H) 7.79 (t, J = 5.28 Hz, 1 H) 7.91 (d, J= 1.54 Hz, 1 H) C, 0.66 see experimental section 24

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.92 (t, J = 7.32 Hz, 3 H) 1.25- 1.44(m, 2 H) 1.60 (quin, J = 7.23 Hz, 2 H) 3.38-3.50 (m, 2 H) 4.49 (d, J =5.12 Hz, 2 H) 5.14 (t, J = 5.49 Hz, 1 H) 5.92 (s, 2 H) 7.14 (d, J = 8.42Hz, 1 H) 7.43 (d, J = 8.05 Hz, 1 H) 7.74 (t, J = 4.76 Hz, 1 H) 7.90 (s,1 H) C, 0.56 see experimental section 25

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89 (t, J = 7.28 Hz, 3 H) 1.17- 1.29(m, 3 H) 1.29-1.39 (m, 2 H) 1.54-1.71 (m, 2 H) 1.76-1.86 (m, 2 H) 2.71(q, J = 7.61 Hz, 2 H) 3.46 (t, J = 6.65 Hz, 2 H) 4.54- 4.63 (m, 1 H)7.36-7.40 (m, 1 H) 7.66 (dd, J = 8.41, 1.63 Hz, 1 H) 7.81 (br. s., 2 H)8.21 (s, 1 H) 8.87 (d, J = 8.53 Hz, 1 H) 12.31 (s 1 H) C, 0.81 seeexperimental section 26

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.90 (t, J = 7.32 Hz, 3 H) 1.27 (d, J =6.95 Hz, 6 H) 1.29-1.40 (m, 2 H) 1.57-1.74 (m, 2 H) 1.74- 1.90 (m, 2 H)2.93-3.05 (m, 1 H) 3.41-3.53 (m, 2 H) 4.54- 4.65 (m, 1 H) 7.38 (d, J =8.42 Hz, 1 H) 7.70 (dd, J = 8.60, 1.65 Hz, 1 H) 8.27 (s, 1 H) 8.98 (d, J= 8.42 Hz, 1 H) 12.49 (s, 1 H) C, 0.85 see experimental section 27

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.87 (t, J = 7.4 Hz, 3 H), 1.23- 1.38(m, 2 H), 1.49-1.62 (m, 2 H), 1.63-1.79 (m, 2 H), 3.44 (t, J = 6.4 Hz, 2H), 4.33-64.42 (m, 1 H), 4.42-4.52 (m, 1 H), 6.43 (br. s., 2 H), 6.99(d, J = 8.8 Hz, 1 H), 7.34 (d, J = 9.0 Hz, 1 H), 7.41 (dd, J = 9.0, 2.5Hz, 1 H), 7.58- 7.68 (m, 2 H), 8.02 (d, J = 2.0 Hz, 1 H), 8.06 (d, J =2.5 Hz, 1 H) C, 1.1 see experimental section 28

¹H NMR (400 MHz, d-DMF) δ ppm 1.36 (t, J = 7.4 Hz, 3 H), 1.79 (dq, J =14.9, 7.4 Hz, 2 H), 1.97- 2.07 (m, 2 H), 3.88 (td, J = 7.0, 5.8 Hz, 2H), 4.74-4.80 (m, 2 H), 4.86-4.92 (m, 2 H), 6.38 (s, 2 H), 7.25 (s, 1H), 8.07 (t, J = 5.5 Hz, 1 H) C, 0.85 see experimental section 29

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.87 (t, J = 7.4 Hz, 3 H), 1.22- 1.39(m, 2 H), 1.46-1.61 (m, 2 H), 1.61-1.79 (m, 2 H), 3.43 (t, J = 6.5 Hz, 2H), 2.48-64.50 (m, 2 H), 6.07 (s, 2 H), 7.10 (d, J = 8.8 Hz, 2 H),7.24-7.40 (m, 3 H), 7.71 (d, J = 8.5 Hz, 2 H), 7.98 (d, J = 2.3 Hz, 1 H)C, 1.05 see experimental section 30

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.28 Hz, 3 H) 1.35- 1.45(m, 2 H) 1.56-1.65 (m, 2 H) 3.44-3.53 (m, 2 H) 3.73 (t, J = 2.38 Hz, 1H) 5.01 (d, J = 2.26 Hz, 2 H) 6.38 (br. s., 2 H) 6.69 (d, J = 8.03 Hz, 1H) 6.86 (d, J = 7.78 Hz, 1 H) 7.42 (t, J = 8.28 Hz, 1 H) 8.04 (br. s., 1H) C, 0.84 Same method as to prepare 11 31

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.40 Hz, 3 H) 1.38 (d, J =6.02 Hz, 6 H) 1.40-1.47 (m, 2 H) 1.56-1.64 (m, 2 H) 3.43- 3.49 (m, 2 H)4.79-4.85 (m, 1 H) 6.08 (br. s., 2 H) 6.61 (d, J = 8.03 Hz, 1 H) 6.76(dd, J = 8.28, 0.75 Hz, 1 H) 7.35 (t, J = 8.16 Hz, 1 H) 7.97 (br. s., 1H) C, 0.96 Same method as to prepare 11 32

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89 (t, J = 7.40 Hz, 3 H) 1.36 (dq, J =14.90, 7.41 Hz, 2 H) 1.56- 1.66 (m, 2 H) 2.82-2.93 (m, 2 H) 3.34-3.43(m, 2 H) 3.43-3.52 (m, 2 H) 5.95 (s, 2 H) 6.60 (t, J = 5.14 Hz, 1 H)6.83-6.89 (m, 1 H) 7.07 (dd, J = 8.28, 1.25 Hz, 1 H) 7.16-7.35 (m, 6 H)C, 1.1 Same method as to prepare 12 33

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89 (t, J = 7.4 Hz, 3 H), 1.21- 1.45(m, 2 H), 1.48-1.71 (m, 2 H), 3.49 (qd, J = 10.4, 5.8 Hz, 2 H),4.31-4.43 (m, 1 H), 64.54 (s, 2 H), 4.71 (br. s., 1 H), 5.27 (br. s., 1H), 6.26 (br. s., 2 H), 7.00 (dd, J = 8.4, 1.4 Hz, 1 H), 7.16 (s, 1 H),7.40 (d, J = 8.0 Hz, 1 H), 8.03 (d, J = 8.5 Hz, 1 H) OH C, 0.51 Samemethod as to prepare 24 34

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89 (t, J = 7.15 Hz, 3 H), 1.34 (td, J= 14.81, 7.78 Hz, 2 H) 1.48- 1.74 (m, 2 H) 3.48 (m, J = 11.70, 5.40 Hz,2 H) 4.38 (m, J = 4.00 Hz, 1 H) 4.50 (d, J = 4.02 Hz, 2 H) 4.68 (t, J =1.00 Hz, 1 H) 5.12 (t, J = 1.00 Hz, 1 H) 5.87 (br. s., 2 H) 7.15 (d, J =8.53 Hz, 1 H) 7.26 (d, J = 8.03 Hz, 1 H) 7.44 (dd, J = 8.50 Hz, 1 H)7.98 (br. s., 1 H) B, 3.04 Same method as to prepare 24 35

¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 0.95 (t, J = 7.4 Hz, 3 H), 1.42(dq, J = 15.1, 7.4 Hz, 2 H), 1.58-1.70 (m, 2 H), 3.56 (td, J = 7.2, 5.6Hz, 2 H), 4.96 (s, 2 H), 5.70 (t, J = 4.8 Hz, 1 H), 6.87 (d, J = 8.5 Hz,1 H), 7.25- 7.30 (m, 2 H), 7.38 (dd, J = 8.5, 1.5 Hz, 1 H), 7.43-7.48(m, 1 H), 7.70 (d, J = 2.0 Hz, 1 H) C, 1.15 Same method as to prepare 2736

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.40 Hz, 3 H) 1.30- 1.47(m, 2 H) 1.55-1.70 (m, 2 H), 2.72 (s, 3 H) 3.42-3.53 (m, 2 H) 5.95 (s, 2H) 6.44-6.60 (m, 1 H) 6.78 (d, J = 7.03 Hz, 1 H) 7.04 (d, J = 7.78 Hz, 1H) 7.29 (dd, J = 8.28, 7.28 Hz, 1 H) C, 0.76 Same method as to prepare 937

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.6 Hz, 3 H), 1.36 (dq, J =14.9, 7.4 Hz, 2 H), 1.55- 1.66 (m, 2 H), 3.42-3.51 (m, 2 H), 6.24 (br.s., 2 H), 66.94 (td, J = 7.9, 5.0 Hz, 1 H), 7.29 (ddd, J = 11.4, 7.8,1.1 Hz, 1 H), 7.79 (d, J = 8.3 Hz, 1 H), 7.84 (t, J = 5.3 Hz, 1 H) C,0.75 Same method as to prepare 9 38

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.4 Hz, 3 H), 1.37 (dq, J =14.9, 7.4 Hz, 2 H), 1.56- 1.67 (m, 2 H), 3.43-3.51 (m, 2 H), 6.38 (br.s., 2 H), 67.26 (dd, J = 9.0, 5.3 Hz, 1 H), 7.42 (td, J = 8.8, 3.0 Hz, 1H), 7.93 (dd, J = 10.2, 2.9 Hz, 1 H), 8.00 (t, J = 5.0 Hz, 1 H) C, 0.76Same method as to prepare 9 39

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.37 Hz, 3 H) 1.28-1.45 (m,2 H) 1.50-1.80 (m, 2 H) 3.40-3.53 (m, 2 H) 3.80 (s, 3 H) 6.07 (br. s, 2H) 6.57-6.70 (m, 1 H) 6.64 (s, 1 H) 7.58 (s, 1 H) 7.81-8.04 (m, 1 H) C,0.71 Same method as to prepare 9 40

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.3 Hz, 3 H), 1.38 (dq, J =14.9, 7.4 Hz, 2 H), 1.57- 1.69 (m, 2 H), 3.44-3.51 (m, 2 H), 3.56 (s,3H), 5.87 (s, 2 H), 7.14-67.19 (m, 2 H), 7.50 (s, 1 H), 7.76 (t, J = 5.4Hz, 1 H) C, 0.71 Same method as to prepare 9 41

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.32 Hz, 3 H) 1.29- 1.44(m, 2 H) 1.63 (quin, J = 7.23 Hz, 2 H) 3.47-3.57 (m, 2 H) 6.67 (br. s.,2 H) 7.14 (dd, J = 7.50, 0.91 Hz, 1 H) 7.21 (dd, J = 8.42, 1.10 Hz, 1 H)7.40-7.51 (m, 1 H) 7.88 (br. s., 1 H) B, 5.78 Same method as to prepare9 42

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.3 Hz, 2 H), 1.36 (dq, J =14.9, 7.4 Hz, 2 H), 1.60 (quin, J = 7.3 Hz, 2 H), 3.40-3.48 (m, 2 H),6.15 (s, 2 H), 67.08 (dd, J = 12.5, 7.8 Hz, 1 H), 7.71 (t, J = 5.3 Hz, 1H), 8.10 (dd, J = 12.0, 9.0 Hz, 1 H) C, 0.87 Same method as to prepare 943

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.78-0.95 (m, 3 H), 1.15- 1.42 (m, 2 H),1.47-1.74 (m, 3 H), 2.37 (s, 3 H), 3.22-3.27 (m, 1 H), 3.42-3.60 (m, 2H), 4.37 (d, J = 5.3 Hz, 1 H), 4.68 (br. s., 1 H), 6.89 (t, J = 7.5 Hz,1 H), 7.18 (d, J = 8.3 Hz, 1 H), 7.33 (d, J = 7.0 Hz, 1 H), 7.89 (d, J =8.0 Hz, 1 H). LC-MS m/z = 261 (M + H) C, 0.64 Same method as to prepare9 44

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89 (t, J = 7.3 Hz, 3 H), 1.20- 1.44(m, 2 H), 1.55 (td, J = 9.1, 4.4 Hz, 1 H), 1.61-1.71 (m, 1 H), 2.33 (s,3 H), 3.41-3.57 (m, 2 H), 4.24-4.43 (m, 1 H), 4.71 (br. s., 1 H), 5.88(s, 2 H), 6.84 (dd, J = 8.3, 1.3 Hz, 1 H), 6.98 (s, 1 H), 7.19 (d, J =8.3 Hz, 1 H), 7.94 (d, J = 8.3 Hz, 1 H) supports structure. LC-MS m/z =261 (M + H) C, 0.64 Same method as to prepare 9 45

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.80-0.92 (m, 3 H) 1.22- 1.43 (m, 2 H)1.48-1.70 (m, 2 H) 2.34 (s, 3 H) 3.47 (ddt, J = 16.81, 10.98, 5.43, 5.43Hz, 2 H) 4.30- 4.40 (m, 1 H) 4.66 (t, J = 5.40 Hz, 1 H) 5.79 (s, 2 H)7.09 (d, J = 8.28 Hz, 1 H) 7.15 (d, J = 8.28 Hz, 1 H) 7.30 (dd, J =8.53, 1.76 Hz, 1 H) 7.86 (s, 1 H) wembrech_1457_2 C, 0.65 Same method asto prepare 9 46

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.85-0.94 (m, 3 H) 1.31- 1.45 (m, 2 H)1.53-1.68 (m, 2 H) 1.90 (s, 3 H) 2.73 (s, 3 H) 3.51- 3.56 (m, 2 H)4.30-4.39 (m, 1 H) 6.00 (s, 2 H) 6.28 (d, J = 8.03 Hz, 1 H) 6.81 (d, J =7.03 Hz, 1 H) 7.05 (d, J = 8.28 Hz, 1 H) 7.30 (t, J = 8.00 Hz, 1 H)wembrech_1405_2 C, 0.66 Same method as to prepare 9 47

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89 (t, J = 7.4 Hz, 3 H), 1.20- 1.45(m, 2 H), 1.47-1.72 (m, 2 H), 3.41-3.56 (m, 2 H), 4.31- 4.43 (m, 1 H),4.69 (br. 6 s., 1 H), 6.24 (br. s., 2 H), 6.95 (td, J = 7.9, 5.0 Hz, 1H), 7.31 (dd, J = 11.3, 7.8 Hz, 1 H), 7.41 (d, J = 8.3 Hz, 1 H), 7.90(d, J = 8.3 Hz, 1 H) C, 0.64 Same method as to prepare 9 48

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.90 (t, J = 7.3 Hz, 3 H), 1.20- 1.45(m, 2 H), 1.51-1.73 (m, 2 H), 3.54 (br. s., 2 H), 4.45 (td, J = 8.5, 5.5Hz, 1 H), 4.82 (br. s., 1 H), 7.18 (dd, J = 10.0, 2.5 Hz, 1 H), 7.25(td, J = 8.8, 2.5 Hz, 1 H), 7.63 (br. s., 2 H), 8.41 (dd, J = 9.0, 5.8Hz, 1 H), 8.60 (d, J = 8.3 Hz, 1 H) C, 0.65 Same method as to prepare 949

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.90 (t, J = 7.4 Hz, 3 H), 1.22- 1.45(m, 2 H), 1.49-1.72 (m, 2 H), 3.43-3.55 (m, 2 H), 4.36 (td, J = 8.7, 5.0Hz, 1 H), 64.69 (br. s., 1 H), 5.98 (s, 2 H), 7.22 (dd, J = 9.0, 5.5 Hz,1 H), 7.27 (d, J = 8.3 Hz, 1 H), 7.37 (td, J = 8.8, 2.8 Hz, 1 H), 7.98(dd, J = 10.3, 2.8 Hz, 1 H) C, 0.63 Same method as to prepare 9 50

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.92 (t, J = 7.4 Hz, 3 H), 1.26- 1.42(m, 2 H), 1.59-1.70 (m, 2 H), 3.53-3.67 (m, 3 H), 4.47 (d, J = 5.3 Hz, 1H), 7.21-7.36 (m, 2 H), 7.80 (td, J = 8.3, 6.0 Hz, 1 H), 7.93 (dd, J =14.8, 8.5 Hz, 1 H), 8.38 (br. s., 1 H), 13.06 (br. s., 1 H). LC-MS m/z =265 (M + H) C, 0.75 Same method as to prepare 9 51

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.79-0.92 (m, 3 H), 1.19- 1.39 (m, 4 H)1.55-1.75 (m, 2 H) 2.41 (s, 3 H) 3.46-3.61 (m, 2 H) 4.40-4.51 (m, 1 H)7.36 (d, J = 8.53 Hz, 1 H) 7.62 (d, J = 8.28 Hz, 1 H) 7.80 (s, 2 H) 8.29(s, 1 H) 8.87 (d, J = 8.28 Hz, 1 H) 12.51 (s, 1 H) wembrech_1457_1 C,0.73 Same method as to prepare 9 52

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.78-0.90 (m, 3 H) 1.20-1.39 (m, 4 H)1.53-1.70 (m, 2 H) 1.90 (s, 3 H) 2.73 (s, 3 H) 3.50- 3.57 (m, 2 H)4.28-4.36 (m, 1 H) 5.98 (s, 2 H) 6.28 (d, J = 8.28 Hz, 1 H) 6.81 (d, J=7.03 Hz, 1 H) 7.05 (d, J = 7.78 Hz, 1 H) 7.30 (t, J = 8.30 Hz, 1 H) C,0.75 Same method as to prepare 9 53

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89 (t, J = 7.3 Hz, 3 H), 1.23- 1.39(m, 2 H), 1.52-1.71 (m, 2 H), 1.74-1.91 (m, 2 H), 2.43 (s, 3 H), 3.45(t, J = 6.5 Hz, 2 H), 4.48- 4.60 (m, 2 H), 7.18-7.29 (m, 2 H), 7.37-8.21(m, 2 H), 8.35 (d, J = 8.3 Hz, 1 H), 8.99 (d, J = 8.3 Hz, 1 H), 12.78(br. s., 1 H) C, 0.69 Same method as to prepare 9 54

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.00 (s, 1 H) 0.79-0.97 (m, 3 H)1.19-1.39 (m, 2 H) 1.51- 1.74 (m, 2 H) 1.74-1.93 (m, 2 H) 2.40 (s, 3 H)3.41-3.52 (m, 2 H) 4.51-4.63 (m, 1 H) 7.35 (d, J = 8.53 Hz, 1 H)7.57-7.65 (m, 1 H) 7.83 (s, 2 H) 8.25 (s, 1 H) 8.91 (d, J = 8.28 Hz, 1H) 12.57 (s, 1 H) wembrech_1457_4 C, 0.72 Same method as to prepare 9 55

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.85-0.95 (m, 3 H) 1.29- 1.42 (m, 2 H)1.53-1.78 (m, 2 H) 1.79-1.86 (m, 2 H) 2.78 (s, 3 H) 3.50-3.66 (m, 2 H)4.57-4.70 (m, 1 H) 7.21 (d, J = 7.28 Hz, 1 H) 7.29 (d, J = 8.03 Hz, 1 H)7.62 (t, J = 7.91 Hz, 1 H) 7.75 (d, J = 8.03 Hz, 2 H) 7.87 (d, J = 8.03Hz, 1 H) 12.36 (s, 1 H) C, 0.75 Same method as to prepare 9 56

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.88 (t, J = 7.3 Hz, 3 H), 1.18-1.43 (m,2 H), 1.54 (td, J = 9.1, 4.4 Hz, 1 H), 1.60-1.71 (m, 1 H), 3.39-3.54 (m,2 H), 3.79 (s, 3 H), 4.33 (td, J = 8.6, 5.1 Hz, 1 H), 4.66 (t, J = 5.4Hz, 1 H), 5.87 (s, 2 H), 6.56-6.65 (m, 2 H), 7.11 (d, J = 8.3 Hz, 1 H),7.95 (d, J = 8.8 Hz, 1 H) C, 0.63 Same method as to prepare 9 57

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.90 (t, J = 7.3 Hz, 3 H), 1.25- 1.45(m, 2 H), 1.57 (dtd, J = 13.7, 9.1, 9.1, 5.0 Hz, 1 H), 1.63-1.75 (m, 1H), 3.44-3.55 6 (m, 2 H), 3.81 (s, 3 H), 4.39 (td, J = 8.5, 5.3 Hz, 1H), 4.70 (br. s., 1 H), 5.74 (s, 2 H), 7.11-7.17 (m, 2 H), 7.23 (d, J =8.3 Hz, 1 H), 7.54 (s, 1 H) C, 0.64 Same method as to prepare 9 58

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.92 (t, J = 7.3 Hz, 3 H), 1.29- 1.43(m, 2 H), 1.56-1.71 (m, 2 H), 3.53-3.65 (m, 2 H), 4.04 (s, 3 H),4.27-4.43 (m, 1 H), 4.66 (br. s., 3 H), 7.02 (d, J = 8.3 Hz, 2 H), 7.71(t, J = 8.3 Hz, 1 H), 8.90 (d, J = 8.3 Hz, 1 H), 12.85 (s, 1 H) C, 0.66Same method as to prepare 9 59

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.85 (t, J = 6.5 Hz, 3 H), 1.19- 1.39(m, 4 H), 1.48-1.62 (m, 1 H), 1.62-1.77 (m, 1 H), 3.40- 3.56 (m, 2 H),4.35 (td, 6 J = 8.7, 5.0 Hz, 1 H), 4.69 (t, J = 5.4 Hz, 1 H), 6.24 (br.s., 2 H), 6.95 (td, J = 8.0, 5.0 Hz, 1 H), 7.31 (dd, J = 11.2, 7.7 Hz, 1H), 7.41 (d, J = 8.3 Hz, 1 H), 7.90 (d, J = 8.3 Hz, 1 H) C, 0.74 Samemethod as to prepare 9 60

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.86 (t, J = 6.7 Hz, 3 H), 1.20- 1.39(m, 4 H), 1.54-1.76 (m, 2 H), 3.55 (d, J = 5.8 Hz, 4 H), 4.37- 4.50 (m,1 H), 7.26 (dd, J = 9.8, 2.5 Hz, 1 H), 7.30-7.36 (m, 1 H), 8.50-8.57 (m,1 H), 8.99 (d, J = 8.0 Hz, 1 H), 12.48 (br. s., 1 H) C, 0.77 Same methodas to prepare 9 61

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.86 (t, J = 6.5 Hz, 3 H), 1.17- 1.40(m, 4 H), 1.47-1.62 (m, 1 H), 1.62-1.76 (m, 1 H), 3.42- 3.55 (m, 2 H),4.25-64.42 (m, 1 H), 4.69 (br. s., 1 H), 6.13 (br. s., 2 H), 7.23 (dd, J= 9.2, 5.4 Hz, 1 H), 7.39 (br. s, 1 H), 7.39 (td, J = 8.6, 2.4 Hz, 1 H),8.00 (dd, J = 10.3, 2.8 Hz, 1 H) C, 0.73 Same method as to prepare 9 62

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.82-0.92 (m, 3 H), 1.25- 1.40 (m, 4 H),1.53-1.73 (m, 2 H), 3.51-3.60 (m, 2 H), 4.37 (m, J = 3.5 Hz, 1 H), 4.92(br. s, 1 H), 6.74 (br. s., 2 H), 6.92 (dd, J = 12.8, 8.0 Hz, 1 H),7.01-7.08 (m, 1 H), 7.08-7.12 (m, 1 H), 7.54 (td, J = 8.2, 6.5 Hz, 1 H).LC- MS m/z = 279 (M + H). C, 0.83 Same method as to prepare 9 63

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.88 (t, J = 7.4 Hz, 3 H), 1.22- 1.40(m, 2 H), 1.47-1.66 (m, 2 H), 1.66-1.80 (m, 2 H), 3.41- 3.49 (m, 2 H),4.33-64.52 (m, 2 H), 6.26 (br. s., 2 H), 6.95 (td, J = 8.0, 4.9 Hz, 1H), 7.31 (dd, J = 11.3, 7.8 Hz, 1 H), 7.48 (d, J = 8.5 Hz, 1 H), 7.87(d, J = 8.3 Hz, 1 H) C, 0.69 Same method as to prepare 9 64

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.88 (t, J = 7.4 Hz, 3 H), 1.21- 1.41(m, 2 H), 1.48-1.66 (m, 2 H), 1.68-1.81 (m, 2 H), 3.42- 3.48 (m, 2 H),4.30-4.55 (m, 2 H), 6.69 (br. s., 2 H), 6.89-7.07 (m, 2 H), 7.86 (d, J =8.3 Hz, 1 H), 8.21 (dd, J = 8.9, 6.1 Hz, 1 H) C, 0.73 Same method as toprepare 9 65

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.88 (t, J = 7.4 Hz, 3 H), 1.25- 1.40(m, 2 H), 1.50-1.65 (m, 2 H), 1.65-1.81 (m, 2 H), 3.45 (t, J = 6.5 Hz, 2H), 4.32-64.52 (m, 2 H), 6.00 (s, 2 H), 7.22 (dd, J = 9.0, 5.5 Hz, 1 H),7.28-7.42 (m, 2 H), 7.95 (dd, J = 10.2, 2.9 Hz, 1 H) C, 0.68 Same methodas to prepare 9 66

¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 0.95 (t, J = 7.3 Hz, 3 H),1.34-1.58 (m, 4 H), 1.59-1.72 (m, 2 H), 1.92- 2.07 (m, 1 H), 3.55-3.73(m, 2 H), 4.42-4.59 (m, 1 H), 5.10 (br. s., 2 H), 6.62 (dd, J = 18.7,8.4 Hz, 1 H), 6.81 (dd, J = 13.1, 8.0 Hz, 1 H), 7.21 (d, J = 8.5 Hz, 1H), 7.42- 7.55 (m, 1 H). LC-MS m/z = 279 (M + H) C, 0.79 Same method asto prepare 9 67

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.86 (t, J = 7.5 Hz, 3 H), 0.93 (d, J =6.8 Hz, 3 H), 1.08-1.24 (m, 1 H), 1.43-1.59 (m, 1 H), 1.84 (ddt, J =11.2, 7.7, 4.0, 64.0 Hz, 1 H), 3.54-3.68 (m, 2 H), 4.20- 4.30 (m, 1 H),4.56 (t, J = 5.4 Hz, 1 H), 6.20 (br. s., 2 H), 6.95 (td, J = 8.0, 5.0Hz, 1 H), 7.30 (ddd, J = 11.4, 7.7, 0.8 Hz, 1 H), 7.39 (d, J = 8.5 Hz, 1H), 7.95 (d, J = 8.3 Hz, 1 H) C, 0.72 Same method as to prepare 9 68

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.86 (t, J = 7.4 Hz, 3 H), 0.92 (d, J =6.8 Hz, 3 H), 1.11-1.24 (m, 1 H), 1.44-1.59 (m, 1 H), 1.83 (ddt, J =11.3, 7.7, 3.9, 63.9 Hz, 1 H), 3.53-3.69 (m, 2 H), 4.16- 4.28 (m, 1 H),4.55 (br. s., 1 H), 5.94 (s, 2 H), 7.21 (dd, J = 9.2, 5.4 Hz, 1 H), 7.28(d, J = 8.3 Hz, 1 H), 7.37 (td, J = 8.8, 2.8 Hz, 1 H), 8.04 (dd, J =10.3, 2.8 Hz, 1 H) F 6 C, 0.71 Same method as to prepare 9 69

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.88 (t, J = 7.28 Hz, 3 H) 1.20- 1.43(m, 2 H) 1.49-1.70 (m, 2 H) 3.40-3.54 (m, 2 H) 4.30- 4.42 (m, 1 H) 4.68(t, J = 5.02 Hz, 1 H) 6.25 (br. s., 2 H) 6.96 (t, J = 7.91 Hz, 1 H) 7.41(d, J = 8.28 Hz, 1 H) 7.62 (d, J = 7.53 Hz, 1 H) 8.04 (d, J = 8.28 Hz, 1H) C, 0.71 Same method as to prepare 9 70

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.80-0.95 (m, 3 H), 1.16- 1.43 (m, 2 H),1.46-1.74 (m, 2 H), 1.91 (t, J = 5.8 Hz, 0 H), 3.43- 3.60 (m, 2 H),3.50-3.50 (m, 0 H), 4.35 (td, J = 8.4, 5.3 Hz, 1 H), 4.79 (br. s., 1 H),6.17 (br. s., 2 H), 7.00 (dd, J = 8.8, 2.0 Hz, 1 H), 7.16 (d, J = 2.0Hz, 1 H), 7.54 (d, J = 8.0 Hz, 1 H), 8.18 (d, J = 8.8 Hz, 1 H). LC-MSm/z = 281 (M + H) supports structure. LC-MS m/z = 281 (M + H) C, 0.72Same method as to prepare 9 71

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.91 (t, J = 7.28 Hz, 3 H) 1.23- 1.44(m, 2 H) 1.52-1.68 (m, 2 H) 3.54 (t, J = 4.14 Hz, 2 H) 4.33 (ddt, J =10.60, 7.22, 3.76, 3.76 Hz, 1 H) 4.90 (t, J = 5.14 Hz, 1 H) 6.22 (br.s., 2 H) 7.05 (dd, J = 7.65, 1.13 Hz, 1 H) 7.15 (dd, J = 8.41, 1.13 Hz,1 H) 7.33-7.42 (m, 1 H) 7.60 (d, J = 8.03 Hz, 1 H) B, 4.98 Same methodas to prepare 9 72

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89 (t, J = 7.3 Hz, 3 H), 1.22- 1.44(m, 2 H), 1.47-1.59 (m, 1 H), 1.59-1.72 (m, 1 H), 3.41- 3.53 (m, 2 H),4.28-64.40 (m, 1 H), 4.68 (t, J = 5.4 Hz, 1 H), 6.11 (s, 2 H), 7.07 (dd,J = 12.5, 7.8 Hz, 1 H), 7.29 (d, J = 8.3 Hz, 1 H), 8.22 (dd, J = 12.0,9.0 Hz, 1 H) C, 0.74 Same method as to prepare 9 73

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.85 (t, J = 6.8 Hz, 3 H), 1.19- 1.40(m, 4 H), 1.56-1.72 (m, 2 H), 1.74-1.92 (m, 2 H), 2.44 (s, 3 H),2.49-2.55 (m, 1 H), 3.46 (t, J = 6.5 Hz, 2 H), 4.47-4.63 (m, 1 H),7.19-7.28 (m, 2 H), 7.92 (d, J = 8.5 Hz, 2 H), 8.37 (d, J = 8.3 Hz, 1H), 9.01 (d, J = 8.3 Hz, 1 H), 12.80 (s, 1 H) C, 0.77 Same method as toprepare 9 74

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.82-0.90 (m, 3 H) 1.22- 1.37 (m, 4 H)1.60-1.68 (m, 2 H) 1.75-1.83 (m, 2 H) 2.42 (s, 3 H) 3.43-3.48 (m, 2 H)4.51-4.59 (m, 1 H) 7.36 (d, J = 8.53 Hz, 1 H) 7.62 (d, J = 8.53 Hz, 1 H)7.74 (br. s., 2 H) 8.19 (s, 1 H) 8.84 (d, J = 8.28 Hz, 1 H) 12.27 (s, 1H) wembrech_1457_3 C, 0.75 Same method as to prepare 9 75

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.82-0.91 (m, 3 H) 1.28- 1.40 (m, 4 H)1.59-1.77 (m, 2 H) 1.83 (q, J = 5.94 Hz, 2 H) 2.78 (s, 3 H) 3.50-3.66(m, 2 H) 4.55- 4.66 (m, 1 H) 7.21 (d, J = 7.53 Hz, 1 H) 7.29 (d, J =8.28 Hz, 1 H) 7.62 (t, J = 7.91 Hz, 1 H) 7.77 (br. s., 2 H) 7.88 (d, J =8.03 Hz, 1 H) 12.38 (s, 1 H) C, 0.83 Same method as to prepare 9 76

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.85 (t, J = 6.40 Hz, 3 H) 1.17- 1.38(m, 4 H) 1.45-1.58 (m, 1 H) 1.62-1.73 (m, 1 H) 3.37- 3.52 (m, 2 H) 3.79(s, 3 H) 4.30 (dd, J = 8.53, 5.02 Hz, 1 H) 4.60- 4.68 (m, 1 H) 5.87 (s,2 H) 6.59- 6.60 (m, 1 H) 6.60-6.65 (m, 1 H) 7.12 (d, J = 8.28 Hz, 1 H)7.96 (d, J = 8.78 Hz, 1 H) wembrech_1505_1 C, 0.72 Same method as toprepare 9 77

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.86 (t, J = 6.5 Hz, 3 H), 1.22- 1.40(m, 4 H), 1.49-1.63 (m, 1 H), 1.65-1.80 (m, 1 H), 3.44- 3.56 (m, 2 H),3.81 (s, 36 H), 4.37 (td, J = 8.5, 5.3 Hz, 1 H), 4.70 (br. s., 1 H),5.73 (s, 2 H), 7.12- 7.17 (m, 2 H), 7.23 (d, J = 8.3 Hz, 1 H), 7.54 (s,1 H) C, 0.73 Same method as to prepare 9 78

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.87 (t, J = 7.40 Hz, 3 H) 1.22- 1.39(m, 2 H) 1.48-1.78 (m, 4 H) 3.37-3.50 (m, 2 H) 3.78 (s, 3 H) 4.34-4.49(m, 1 H) 4.34- 4.49 (m, 1 H) 5.92 (s, 2 H) 6.60 (d, J = 2.51 Hz, 1 H)6.61-6.66 (m, 1 H) 7.21 (d, J = 8.53 Hz, 1 H) 7.94 (d, J = 8.78 Hz, 1 H)wembrech_1505_4 C, 0.67 Same method as to prepare 9 79

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89 (t, J = 7.3 Hz, 3 H), 1.26- 1.41(m, 2 H), 1.51-1.66 (m, 2 H), 1.66-1.83 (m, 2 H), 3.42- 3.47 (m, 1 H),3.81 (s, 3 H), 4.38- 4.52 (m, 2 H), 5.87 (s, 2 H), 7.14-7.19 (m, 2 H),7.35 (d, J = 8.5 Hz, 1 H), 7.53 (s, 1 H) C, 0.67 Same method as toprepare 9 80

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.86 (t, J = 7.4 Hz, 3 H), 0.94 (d, J =6.4 Hz, 3 H), 1.11-1.24 (m, 1 H), 1.53 (ddd, J = 13.4, 7.5, 3.9 Hz, 1H), 1.87 (ddt, 6 J = 11.2, 7.7, 4.0, 4.0 Hz, 1 H), 3.58-3.66 (m, 2 H),3.82 (s, 3 H), 4.20-4.31 (m, 1 H), 4.58 (br. s., 1 H), 5.69 (s, 2 H),7.12-7.17 (m, 2 H), 7.24 (d, J = 8.5 Hz, 1 H), 7.59 (s, 1 H) C, 0.7 Samemethod as to prepare 9 81

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.84 (t, J = 6.5 Hz, 3 H), 1.20- 1.37(m, 4 H), 1.52-1.65 (m, 2 H), 1.65-1.80 (m, 2 H), 3.44 (q, J = 6.2 Hz, 2H), 4.35-64.49 (m, 2 H), 6.25 (br. s., 2 H), 6.95 (td, J = 7.9, 5.0 Hz,1 H), 7.31 (dd, J = 11.3, 7.8 Hz, 1 H), 7.48 (d, J = 8.3 Hz, 1 H), 7.87(d, J = 8.3 Hz, 1 H) C, 0.79 Same method as to prepare 9 82

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.79-0.89 (m, 3 H), 1.19- 1.37 (m, 4 H),1.59 (d, J = 6.5 Hz, 2 H), 1.65-1.79 (m, 2 H), 3.43 (t, J = 6.3 Hz, 2H), 4.31-4.53 (m, 2 H), 6.24 (s, 2 H), 6.80-6.98 (m, 2 H), 7.51 (d, J =8.5 Hz, 1 H), 8.14 (dd, J = 8.8, 6.5 Hz, 1 H) C, 0.81 Same method as toprepare 9 83

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.85 (t, J = 6.3 Hz, 3 H), 1.20- 1.37(m, 4 H), 1.53-1.64 (m, 2 H), 1.64-1.82 (m, 2 H), 3.45 (t, J = 6.4 Hz, 2H), 4.34-64.48 (m, 2 H), 6.01 (s, 2 H), 7.22 (dd, J = 9.2, 5.4 Hz, 1 H),7.29-7.42 (m, 2 H), 7.95 (dd, J = 10.3, 2.8 Hz, 1 H) C, 0.77 Same methodas to prepare 9 84

¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 0.89 (t, J =7.0 Hz, 3 H), 1.19-1.46(m, 4 H), 1.50-1.79 (m, 4 H), 1.92- 2.12 (m, 1 H), 3.59-3.75 (m, 2 H),3.96 (br. s., 2 H), 4.40-4.56 (m, 1 H), 6.72 (dd, J = 18.6, 8.5 Hz, 1H), 6.81 (ddd, J = 12.8, 8.0, 0.8 Hz, 1 H), 7.19 (d, J = 8.5 Hz, 1 H),7.48 (td, J = 8.2, 6.4 Hz, 1 H). LC-MS m/z = 2.93 (M + H) C, 0.88 Samemethod as to prepare 9 85

¹H NMR (400 MHz, METHANOL- d₄) δ ppm 0.99 (t, J = 7.3 Hz, 3 H),1.38-1.50 (m, 2 H), 1.71 (quin, J = 7.4 Hz, 2 H), 3.66 (t, J = 7.3 Hz, 2H), 7.33 (d, J = 8.8 Hz, 1 H), 7.87 (dd, J = 8.8, 1.8 Hz, 1 H), 8.00(br. s., 1 H), 8.35 (d, J = 2.0 Hz, 1 H), exchangeable protons not seenC, 0.9 Same method as to prepare 9 86

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89 (t, J = 7.3 Hz, 3 H), 1.24- 1.44(m, 2 H), 1.50-1.73 (m, 2 H), 3.50 (tq, J = 11.1, 5.3 Hz, 2 H), 3.88 (s,3 H), 4.386 (td, J = 8.6, 5.1 Hz, 1 H), 4.69 (t, J = 5.1 Hz, 1 H), 6.17(br. s., 2 H), 7.50 (dd, J = 8.5, 1.8 Hz, 2 H), 7.74 (d, J = 1.8 Hz, 1H), 8.19 (d, J = 8.5 Hz, 1 H) C, 0.68 Same method as to prepare 9 87

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.76-0.89 (m, 3 H) 1.28 (d, J = 5.02 Hz,4 H) 1.48-1.78 (m, 4 H) 3.36-3.48 (m, 2 H) 3.69- 3.84 (m, 3 H) 4.32-4.46(m, 1 H) 4.32-4.46 (m, 1 H) 5.90 (s, 2 H) 6.60 (d, J = 2.51 Hz, 1 H)6.63 (s, 1 H) 7.20 (d, J = 8.53 Hz, 1 H) 7.94 (d, J = 9.03 Hz, 1 H) C,0.74 Same method as to prepare 9 88

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.85 (t, J = 6.7 Hz, 2 H), 1.22- 1.36(m, 4 H), 1.56-1.65 (m, 2 H), 1.65-1.84 (m, 2 H), 3.40- 3.50 (m, 2 H),3.81 (s, 36 H), 4.38-4.49 (m, 2 H), 5.74 (s, 2 H), 7.15 (s, 2 H), 7.27(d, J = 8.5 Hz, 1 H), 7.51 (s, 1 H) C, 0.76 Same method as to prepare 989

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.4 Hz, 3 H), 1.31- 1.44(m, 2 H), 1.55-1.66 (m, 2 H), 3.40-3.50 (m, 2 H), 3.80 (s, 3 H), 5.05(s, 2 H), 5.67 (s, 2 H), 6.66 (s, 1 H), 7.32-7.46 (m, 4 H), 7.48-7.50(m, 1 H), 7.51 (m, J = 1.5 Hz, 1 H), 7.59 (s, 1 H) C, 0.94 Same methodas to prepare 9 90

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.4 Hz, 3 H), 1.38 (dq, J =15.0, 7.3 Hz, 2 H), 1.63 (quin, J = 7.4 Hz, 2 H), 3.43-3.54 (m, 2 H),6.19 (s, 2 H), 67.37- 7.44 (m, 1 H), 7.52 (dd, J = 8.4, 1.8 Hz, 1 H),7.82 (d, J = 1.5 Hz, 1 H), 7.93 (t, J = 5.4 Hz, 1 H), 8.12 (d, J = 8.6Hz, 1 H), 8.19-8.25 (m, 1 H), 8.32 (dd, J = 4.7, 1.4 Hz, 1 H), 8.97 (d,J = 2.2 Hz, 1 H), 10.54 (s, 1 H) C, 0.73 See experimental 91

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.4 Hz, 3 H), 1.36 (dq, J =14.9, 7.4 Hz, 2 H), 1.55- 1.66 (m, 2 H), 3.43-3.50 (m, 2 H), 3.50-3.74(m, 4 H), 66.21 (br. s., 2 H), 6.99 (dd, J = 8.3, 1.7 Hz, 1 H), 7.12 (d,J = 1.5 Hz, 1 H), 7.88 (t, J = 5.4 Hz, 1 H), 8.04 (d, J = 8.4 Hz, 1 H)C, 0.65 Same as to prepare 90 92

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.4 Hz, 3 H), 1.37 (dq, J =15.0, 7.4 Hz, 2 H), 1.62 (quin, J = 7.3 Hz, 2 H), 2.19 (s, 6 H), 2.41(t, J = 6.8 Hz, 2 H), 3.30- 3.42 (m, 2 H), 3.42-3.51 (m, 2 H), 6.13 (s,2 H), 7.40 (dd, J = 8.4, 1.8 Hz, 1 H), 7.64 (d, J = 1.8 Hz, 1 H), 7.85(t, J = 5.4 Hz, 1 H), 8.03 (d, J = 8.6 Hz, 1 H), 8.45 (t, J = 5.6 Hz, 1H) C, 0.8 Same as to prepare 90 93

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.4 Hz, 3 H), 1.37 (dq, J =14.9, 7.4 Hz, 2 H), 1.62 (quin, J = 7.3 Hz, 2 H), 3.44-3.52 (m, 2 H),4.58 (d, J = 5.96 Hz, 2 H), 6.16 (br. s., 2 H), 7.27 (dd, J = 7.2, 5.0Hz, 1 H), 7.34 (d, J = 7.9 Hz, 1 H), 7.49 (dd, J = 8.5, 1.7 Hz, 1 H),7.71-7.80 (m, 2 H), 7.88 (t, J = 5.4 Hz, 1 H), 8.07 (d, J = 8.4 Hz, 1H), 8.52 (dd, J = 4.8, 0.7 Hz, 1 H), 9.18 (t, J = 5.9 Hz, 1 H) C, 0.71Same as to prepare 90 94

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.4 Hz, 3 H), 1.37 (dq, J =14.9, 7.4 Hz, 2 H), 1.55- 1.67 (m, 2 H), 2.90 (s, 3 H), 2.99 (s, 3 H),3.43-3.52 (m, 2 H), 6.26 (br. s., 2 H), 6.99 (dd, J = 8.3, 1.7 Hz, 1 H),7.12 (d, J = 1.5 Hz, 1 H), 7.93 (t, J = 5.2 Hz, 1 H), 8.04 (d, J = 8.4Hz, 1 H) C, 0.65 Same as to prepare 90 95

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.93 (t, J = 7.4 Hz, 3 H), 1.31- 1.43(m, 2 H), 1.62 (quin, J = 7.3 Hz, 2 H), 3.43-3.51 (m, 2 H), 4.49 (d, J =5.9 Hz, 2 H), 66.22 (br. s., 2 H), 7.20-7.29 (m, 1 H), 7.30-7.35 (m, 4H), 7.48 (dd, J = 8.4, 1.8 Hz, 1 H), 7.72 (d, J = 1.8 Hz, 1 H), 7.93 (t,J = 5.3 Hz, 1 H), 8.07 (d, J = 8.6 Hz, 1 H), 9.13 (t, J = 5.9 Hz, 1 H)C, 0.87 Same as to prepare 90 96

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.90 (t, J = 7.37 Hz, 3 H) 1.31- 1.41(m, 2 H) 1.57-1.65 (m, 2 H) 2.78-2.84 (m, 2 H) 3.36-3.37 (m, 2 H)3.43-3.51 (m, 2 H) 3.72 (s, 3 H) 6.03 (br. s., 2 H) 6.63 (br. s., 1 H)6.82-6.88 (m, 3 H) 7.05-7.14 (m, 3 H) 7.30-7.34 (m, 1 H) C, 1.08 Same asto prepare 12 97

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89 (t, J = 7.37 Hz, 3 H) 1.32- 1.39(m, 2 H) 1.58-1.64 (m, 2 H) 2.82-2.87 (m, 2 H) 3.35-3.41 (m, 2 H)3.45-3.51 (m, 2 H) 3.72 (s, 3 H) 6.03 (br. s., 2 H) 6.67 (br. s., 1 H)6.74- 6.80 (m, 3 H) 6.89 (d, J = 7.04 Hz, 1 H) 7.08 (d, J = 7.26 Hz, 1H) 7.19 (t, J = 8.03 Hz, 1 H) 7.33 (t, J = 7.70 Hz, 1 H) C, 1.06 Same asto prepare 12 98

¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.94 (t, J = 7.37 Hz, 3 H) 1.34- 1.47(m, 2 H) 1.60-1.71 (m, 2 H) 3.42-3.56 (m, 2 H) 3.79 (s, 3 H) 4.93 (s, 2H) 6.03 (s, 2 H) 6.48- 6.57 (m, 1 H) 6.81 (dd, J = 8.36, 0.66 Hz, 1 H)7.33 (t, J = 8.25 Hz, 1 H) 8.25-8.34 (m, 1 H) C, 0.84 Same as to prepare11

Analytical Methods.

All compounds were characterized by LC-MS. The following LC-MS methodswere used:

Method A.

Reversed phase UPLC (Ultra Performance Liquid Chromatography) wascarried out on a bridged ethylsiloxane/silica hybrid (BEH) C18 column(1.7 μm, 2.1×50 mm; Waters Acquity) with a flow rate of 0.8 mL/min. Twomobile phases (10 mM ammonium acetate in H₂O/acetonitrile 95/5; mobilephase B: acetonitrile) were used to run a gradient condition from 95% Aand to 5% B to 5% A and 95% B in 1.3 minutes and hold for 0.7 minutes.An injection volume of 0.75 μl was used. Cone voltage was 30 V forpositive ionization mode and 30 V for negative ionization mode.

Method B.

Reversed phase HPLC was carried out on an Xterra MS C18 column (3.5 μm,4.6×100 mm) with a flow rate of 1.6 mL/min. Three mobile phases (mobilephase A: 95% 25 mM ammoniumacetate+5% acetonitrile; mobile phase B:acetonitrile; mobile phase C: methanol) were employed to run a gradientcondition from 100% A to 50% B and 50% C in 6.5 minutes, to 100% B in0.5 minute, 100% B for 1 minute and reequilibrate with 100% A for 1.5minutes. An injection volume of 10 μl was used.

Method C.

Reversed phase UPLC (Ultra Performance Liquid Chromatography) wascarried out on a bridged ethylsiloxane/silica hybrid (BEH) C18 column(1.7 μm, 2.1×50 mm; Waters Acquity) with a flow rate of 0.8 mL/min. Twomobile phases (mobile phase A: 10 mM ammonium acetate inH₂O/acetonitrile 95/5; mobile phase B: acetonitrile) were used to run agradient condition from 95% A and 5% B to 5% A and 95% B in 1.3 minutesand hold for 0.2 minutes. An injection volume of 0.5 μl was used.

Biological Activity of Compounds of Formula (I) Description ofBiological Assays Assessment of TLR7 and TLR8 Activity

The ability of compounds to activate human TLR7 and/or TLR8 was assessedin a cellular reporter assay using HEK293 cells transiently transfectedwith a TLR7 or TLR8 expression vector and NFκB-luc reporter construct.In one instance the TLR expression construct expresses the respectivewild type sequence or a mutant sequence comprising a deletion in thesecond leucine-rich repeat of the TLR. Such mutant TLR proteins havepreviously been shown to be more susceptible to agonist activation (U.S.Pat. No. 7,498,409).

Briefly, HEK293 cells were grown in culture medium (DMEM supplementedwith 10% FCS and 2 mM Glutamine). For transfection of cells in 10 cmdishes, cells were detached with Trypsin-EDTA, transfected with a mix ofCMV-TLR7 or TLR8 plasmid (750 ng), NFκB-luc plasmid (375 ng) and atransfection reagent and incubated 24 hours at 37° C. in a humidified 5%CO₂ atmosphere. Transfected cells were then detached with Trypsin-EDTA,washed in PBS and resuspended in medium to a density of 1.67×10⁵cells/mL. Thirty microliters of cells were then dispensed into each wellin 384-well plates, where 10 μL of compound in 4% DMSO was alreadypresent. Following 6 hours incubation at 37° C., 5% CO₂, the luciferaseactivity was determined by adding 15 μl of Steady Lite Plus substrate(Perkin Elmer) to each well and readout performed on a ViewLux ultraHTSmicroplate imager (Perkin Elmer). Dose response curves were generatedfrom measurements performed in quadruplicates. Lowest effectiveconcentrations (LEC) values, defined as the concentration that inducesan effect which is at least two fold above the standard deviation of theassay, were determined for each compound.

Compound toxicity was determined in parallel using a similar dilutionseries of compound with 30 μL per well of cells transfected with theCMV-TLR7 construct alone (1.67×10⁵ cells/mL), in 384-well plates. Cellviability was measured after 6 hours incubation at 37° C., 5% CO₂ byadding 15 μL of ATP lite (Perkin Elmer) per well and reading on aViewLux ultraHTS microplate imager (Perkin Elmer). Data was reported asCC₅₀.

Suppression of HCV Replicon Replication

Activation of human TLR7 results in robust production of interferon byplasmacytoid dendritic cells present in human blood. The potential ofcompounds to induce interferon was evaluated by looking at the antiviralactivity in the HCV replicon system upon incubation with conditionedmedia from peripheral blood mononuclear cells (PBMC). The HCV repliconassay is based on a bicistronic expression construct, as described byLohmann et al. (Science (1999) 285: 110-113; Journal of Virology (2003)77: 3007-15 3019) with modifications described by Krieger et al.(Journal of Virology (2001) 75: 4614-4624). The assay utilized thestably transfected cell line Huh-7 luc/neo harboring an RNA encoding abicistronic expression construct comprising the wild type NS3-NS5Bregions of HCV type 1b translated from an Internal Ribosome Entry Site(IRES) from encephalomyocarditis virus (EMCV), preceded by a reportergene (Firefly-luciferase) and a selectable marker gene (neoR, neomycinephosphotransferase). The construct is flanked by 5′ and 3′ NTRs(non-translated regions) from HCV type 1b. Continued culture of thereplicon cells in the presence of G418 (neoR) is dependent on thereplication of the HCV RNA. The stably transfected replicon cells thatreplicate HCV RNA autonomously and to high levels, encoding inter alialuciferase, were used for profiling of the conditioned cell culturemedia.

Briefly, PBMCs were prepared from buffy coats of at least two donorsusing a standard Ficoll centrifugation protocol. Isolated PBMCs wereresuspended in RPMI medium supplemented with 10% human AB serum and2×10⁵ cells/well were dispensed into 384-well plates containingcompounds (70 μL total volume). After overnight incubation, 10 μL ofsupernatant was transferred to 384-well plates containing 2.2×10³replicon cells/well in 30 μL (plated the day before). Following 24 hoursof incubation, replication was measured by assaying luciferase activityusing 40 μL/well Steady Lite Plus substrate (Perkin Elmer) and measuredwith ViewLux ultraHTS microplate imager (Perkin Elmer). The inhibitoryactivity of each compound on the Huh7-luc/neo cells were reported asEC₅₀ values, defined as the compound concentration applied to the PBMCsresulting in a 50% reduction of luciferase activity which in turnindicates the degree of replication of the replicon RNA on transfer of adefined amount of PBMC culture medium. Recombinant interferon α-2a(Roferon-A) was used as a standard control compound.

Biological activity of compounds of formula (I). All compounds showedCC50 of >24 μM in the HEK 293 TOX assay described above.

Activation of ISRE Promoter Elements

The potential of compounds to induce IFN-I was also evaluated bymeasuring the activation of interferon-stimulated responsive elements(ISRE) by conditioned media from PBMC. The ISRE element of sequenceGAAACTGAAACT is highly responsive to the STAT1-STAT2-IRF9 transcriptionfactor, activated upon binding of IFN-I to their receptor IFNAR(Clontech, PT3372-5W). The plasmid pISRE-Luc from Clontech (ref. 631913)contains 5 copies of this ISRE element, followed by the fireflyluciferase ORF. A HEK293 cell line stably transfected with pISRE-Luc(HEK-ISREluc) was established to profile of the conditioned PBMC cellculture media.

Briefly, PBMCs were prepared from buffy coats of at least two donorsusing a standard Ficoll centrifugation protocol. Isolated PBMCs wereresuspended in RPMI medium supplemented with 10% human AB serum and2×10⁵ cells/well were dispensed into 384-well plates containingcompounds (70 μL total volume). After overnight incubation, 10 μL ofsupernatant was transferred to 384-well plates containing 5×10³HEK-ISREluc cells/well in 30 μL (plated the day before). Following 24hours of incubation, activation of the ISRE elements was measured byassaying luciferase activity using 40 μL/well Steady Lite Plus substrate(Perkin Elmer) and measured with ViewLux ultraHTS microplate imager(Perkin Elmer). The stimulating activity of each compound on theHEK-ISREluc cells was reported as LEC value, defined as the compoundconcentration applied to the PBMCs resulting in a luciferase activity atleast two fold above the standard deviation of the assay. The LEC inturn indicates the degree of ISRE activation on transfer of a definedamount of PBMC culture medium. Recombinant interferon α-2a (Roferon-A)was used as a standard control compound.

For a given compound, the LEC value obtained from this assay were in thesame range as the EC₅₀ values obtained from the “suppression of HCVreplication assay.” Thus, it is possible to compare the potential ofcompounds to induce IFN-I by PBMC, measured by either of the 2 assays.

TLR7- TLR7- TLR8- TLR8- PBMC- # STRUCTURE wt_LEC dIRR2_LEC wt_LECdIRR2_LEC HUH7_EC50 1

5.0 0.4 1.1 0.6 1.9 2

NA 1.2 1.5 0.6 4.4 3

 4.0 * 0.9 5.5 2.4 0.6 4

NA 2.4 2.6 1.7 3.0 5

NA 2.6 6.7 2.6 3.3 6

NA 3.4 4.4 2.3 3.0 7

NA 3.8 13.8  9.0 12.4  8

0.1  0.02 0.1  0.02 NA TLR7-wt TLR8-wt HEK-ISRE luc # Structure (LEC)(LEC) (LEC)  9

0.41 0.13 0.10 10

6.08 >25    2.14 11

0.08 0.17 0.12 12

1.66 0.79 NA 13

0.76 0.30 0.57 14

0.65 4.77 NA 15

0.49 3.27 NA 16

0.57 0.73 NA 17

5.10 1.66 0.75 18

0.13 0.13 0.05 19

>25    7.05 NA 20

>25    2.55 NA 21

>25    2.55 12.06  22

6.07 1.95 NA 23

10.23  5.05 NA 24

0.93 0.22 0.14 25

0.57 0.45 0.16 26

3.60 1.97 3.07 27

12.95  >25    14.21  28

2.06 0.88 1.11 29

11.13  >25    >23.81  30

0.05 0.10 0.04 31

0.09 0.24 0.04 32

0.13 0.47 0.27 33

1.88 0.15 0.14 34

10.77  0.27 0.39 35

4.15 >25    >23.81  36

0.47 0.26 0.32 37

0.16 0.25 0.07 38

0.29 0.10 0.11 39

0.94 0.31 0.15 40

5.64 1.83 2.44 41

0.99 0.13 0.17 42

1.81 0.14 0.26 43

4.54 0.12 0.59 44

0.43 0.03 0.09 45

0.41 0.03 0.04 46

0.77 0.04 0.07 47

0.67 0.03 0.05 48

0.54 0.01 0.02 49

2.09 0.03 0.13 50

0.32 0.00 0.01 51

0.60 0.04 0.09 52

0.41 0.03 0.03 53

0.06 0.05 0.02 54

0.54 0.43 0.18 55

0.22 0.14 0.06 56

0.39 0.04 0.09 57

10.77  0.53 2.08 58

0.18 0.03 0.04 59

0.29 0.04 0.05 60

0.23 0.01 0.02 61

0.57 0.05 0.12 62

0.75 0.01 0.03 63

0.29 0.15 0.04 64

0.11 0.03 0.04 65

0.94 0.44 0.56 66

0.22 0.02 0.06 67

2.50 0.11 0.21 68

4.57 0.33 0.64 69

7.48 0.38 0.73 70

0.41 0.01 0.01 71

1.02 0.01 0.03 72

2.59 0.02 0.05 73

0.03 0.06 0.02 74

0.44 0.25 0.14 75

0.14 0.06 0.02 76

0.26 0.04 0.09 77

3.48 0.62 1.93 78

0.20 0.13 0.04 79

11.87  2.97 2.07 80

>25    4.10 >24    81

0.11 0.16 0.05 82

0.04 0.03 0.04 83

1.59 0.42 0.37 84

0.44 0.10 0.09 85

0.51 0.10 0.21 86

2.01 0.22 0.28 87

0.16 0.16 0.04 88

1.85 2.81 0.88 89

1.84 2.22 >24    90

0.42 NA NA 91

0.13 0.53 NA 92

1.53 5.87 NA 93

0.77 1.69 NA 94

0.07 0.48 NA 95

0.54 0.42 NA 96

2.7  16    NA 97

0.6  0.83 NA 98

0.21 0.31 NA * Assay run at 48 hours

1. A compound of formula (I)

or a pharmaceutically acceptable salt, solvate or polymorph thereof,wherein R₁ is C₃₋₈alkyl, C₃₋₈alkoxy, C₂₋₆alkenyl or C₂₋₆alkynyl, each ofwhich is optionally substituted by one or more substituentsindependently selected from halogen, hydroxyl, amino, nitrile, ester,amide, C₁₋₃alkyl, C₁₋₃alkoxy or C₃₋₆cycloalkyl, R₂ is hydrogen, halogen,hydroxyl, amine, C₁₋₇alkyl, C₁₋₇alkylamino, C₁₋₆alkoxy,(C₁₋₄)alkoxy-(C₁₋₄)alkyl, C₃₋₆cycloalkyl, C₄₋₇heterocycle, aromatic,bicyclic heterocycle, arylalkyl, heteroaryl, heteroarylalkyl, carboxylicamide, carboxylic ester each of which is optionally substituted by oneor more substituents independently selected from halogen, hydroxyl,amino, C₁₋₆alkyl, di-(C₁₋₆)-alkylamino, C₁₋₆alkylamino, C₁₋₆alkyl,C₁₋₆alkoxy, C₃₋₆cycloalkyl, carboxylic acid, carboxylic ester,carboxylic amide, heterocycle, aryl, alkenyl, alkynyl, arylalkyl,heteroaryl, heteroarylalkyl, or nitrile, R₃ is hydrogen, halogen,hydroxyl, amine, C₁₋₇alkyl, C₁₋₇alkenyl, C₁₋₇alkynyl, C₁₋₇alkylamino,C₁₋₆alkoxy, (C₁₋₄)alkoxy-(C₁₋₄)alkyl, C₃₋₆cycloalkyl, C₄₋₇hetero-cycle,aromatic, bicyclic heterocycle, arylalkyl, heteroaryl, heteroarylalkyl,aryloxy, heteroaryloxy, ketone, nitrile each of which is optionallysubstituted by one or more substituents independently selected fromhalogen, hydroxyl, amino, C₁₋₆alkyl, di-(C₁₋₆)alkylamino,C₁₋₆alkylamino, C₁₋₆alkyl, C₁₋₆alkoxy, C₃₋₆cycloalkyl, carboxylic acid,carboxylic ester, carboxylic amide, heterocycle, aryl, alkenyl, alkynyl,arylalkyl, heteroaryl, heteroarylalkyl, or nitrile. R₄ is hydrogen,halogen, hydroxyl, amine, C₁₋₇alkyl, C₁₋₇alkylamino, C₁₋₆alkoxy,(C₁₋₄)alkoxy-(C₁₋₄)alkyl, C₃₋₆cycloalkyl, C₄₋₇heterocycle, bicyclicheterocycle, arylalkyl, heteroarylalkyl, aryloxy, heteroaryloxy each ofwhich is optionally substituted by one or more substituentsindependently selected from halogen, hydroxyl, amino, C₁₋₆alkyl,di-(C₁₋₆)alkylamino, C₁₋₆alkylamino, C₁₋₆alkyl, C₁₋₆alkoxy,C₃₋₆cycloalkyl, carboxylic acid, carboxylic ester, carboxylic amide,heterocycle, aryl, alkenyl, alkynyl, arylalkyl, heteroaryl,heteroarylalkyl, or nitrile, and R₅ is hydrogen, fluorine, chlorine ormethyl with the proviso that R₂, R₃, R₄, and R₅ cannot all be H.
 2. Acompound of formula (I) according to claim 1 wherein R₁ is butyl, pentylor 2-pentyl and wherein R₂, R₃, R₄ and R₅ are as specified above.
 3. Acompound of formula (I) according to claim 1 wherein R₁ is C₄₋₈alkylsubstituted with a hydroxyl, and wherein R₂, R₃, R₄ and R₅ are asspecified above.
 4. A compound of formula (I) according to claim 3wherein R₁, when being C₄₋₈alkyl substituted with hydroxyl, is one ofthe following:


5. A compound of formula (I) according to claim 1 wherein R₅ ispreferably hydrogen or fluorine and R₁, R₂, R₃, and R₄ are as describedabove.
 6. A pharmaceutical composition comprising a compound of claim 1or a pharmaceutically acceptable salt, solvate or polymorph thereof andone or more pharmaceutically acceptable excipients, diluents orcarriers.
 7. (canceled)
 8. A method for the treatment of a subjecthaving a disorder in which the modulation of TLR7 and/or TLR8 isinvolved, said method comprising administration of a compound claim 1 ora pharmaceutically acceptable salt, solvate or polymorph thereof, or apharmaceutical composition according to claim 6 to the subject.
 9. Acompound of claim 1 having a structure shown in Table 1, or apharmaceutically acceptable salt, solvate or polymorph thereof.
 10. Acompound of claim 1 having the structure:

or a pharmaceutically acceptable salt, solvate or polymorph thereof.